Therapeutic quinolone compounds with 5-ht-antagonistic properties

ABSTRACT

Provided herein is a compound having the formula (I) (R 2  is typically a tertiary nitrogen atom, being either alkyl-substituted or member of q heterocyclic ring; R 7  is typically a monocyclic or bicyclic aromatic ring or a heterocyclic ring) wherein said compounds are useful for the treatment of psychiatric disorders including but not limited to depression, generalized anxiety, eating disorders, dementia, panic disorder, and sleep disorders. The compounds may also be useful in the treatment of gastrointestinal disorders, cardiovascular regulation, motor disorders, endocrine disorders, vasospasm and sexual dysfunction. The compounds are 5HT 1B  and 5HT 1D  antagonists.

FIELD OF THE INVENTION

This invention relates to novel 8-amino derivatives, methods for theirpreparation, pharmaceutical compositions containing them and their usein therapy.

BACKGROUND OF THE INVENTION

Serotonin (5-HT) has been implicated in many psychiatric disordersincluding but not limited to depression, generalized anxiety, eatingdisorders, dementia, panic disorder, and sleep disorders. Furthermoreserotonin has been implicated in gastrointestinal disorders,cardiovascular regulation, motor disorders, endocrine disorders,vasospasm and sexual dysfunction. Serotonin receptors have beensubdivided into at least 14 subtypes, see Barnes and Sharp,Neuropharmacology, 1999, 38, 1083-1152, incorporated herein byreference. These various subtypes are responsible for serotonin's actionin many pathophysicogical conditions. The 5-HT₁ family of receptors hashigh affinity for serotonin and consists of five related receptors. Thisfamily includes the 5-HT_(1B) and 5-HT_(1D) receptor subtypes. Compoundsthat interact with the 5-HT₁ family are known to have therapeuticpotential in the above mentioned disorders and diseases. In particular,compounds that are 5HT_(1B) and 5HT_(1D) antagonist have been known tobe antidepressant and anxiolytic agents. Compounds that are 5HT_(1B) and5HT_(1D) agonists have been used in the treatment of migraine.

SUMMARY OF THE INVENTION

Provided herein is a compound having the formula (I):

wherein

-   R¹ is, at each position, independently represented by hydrogen,    optionally substituted alkyl, optionally substituted cycloalkyl,    methoxy, thiomethoxy, —NHA, —NA₂, —NHC(═O)A, aminocarbonyl,    —C(═O)NHA, —C(═O)NA₂, halogen, hydroxy, —OA, cyano or aryl;-   A is optionally substituted alkyl, optionally substituted    cycloalkyl, optionally substituted alkenyl or optionally substituted    alkynyl;-   R² is represented by (i), (ii), (iii), or (iv) below:-   R³ is —H, optionally substituted C₁₋₆alkyl, optionally substituted    C₂₋₆alkenyl, optionally substituted C₂₋₆alkynyl, optionally    substituted C₃₋₆cycloalkyl or AOH;-   n is 2, 3 or 4;-   P is a heterocyclic ring;-   R⁴ is —H or optionally substituted C₁₋₄alkyl;-   R⁵ is ═O, ═NR⁴ or ═S;-   R⁶ is —H or methyl;-   Y is —C(═O)NH—, —C(═O)NA-, —C(═O)N(A)-, —NHC(═O)—, —C(═S)NH—,    —CH₂NH—, —C(═O)CH₂—, —CH₂C(═O)—, —C(═O)-piperazine-, —C(═O)R⁸—,    —NAC(═O)—, —C(═S)N(A)-, —CH₂N(A)-, —N(A)CH₂— or a 5-membered    heterocyclic.-   R⁷ is a monocyclic or bicyclic aromatic ring or a heterocycle,    optionally substituted by one or more substituents selected from    R⁸-R⁹ and R¹⁰; wherein R⁷ is connected to Y either by a single bond    or by a ring fusion;-   R⁸ is —CH₂—, —C(═O)—, —SO₂—, —SO₂NH—, —C(═O)NH—, —O—, —S—, —S(═O)—,    a five-membered heterocyclic connected to R⁷ by a ring fusion or a    single bond as tether;-   R⁹ is morpholine optionally substituted with at least one    substituent selected from A, thiomorpholine, piperazin-R¹¹,    optionally substituted aryl, optionally substituted heterocycle, or    —C(═O)CA;-   R¹⁰ is optionally substituted alkyl, optionally substituted    cycloalkyl, hydroxy, aryl, cyano, halogen, —C(═O)NH₂—, methylthio,    —NHA, —NA₂, —NHC(═O)A, —C(═O)NHA, —C(═O)NA₂, or OA;-   R¹¹ is —H, alkyl, AOH, —SO₂A, —SO₂NH₂, —SO₂NHA, —SO₂NA₂, —SO₂NHAR⁹,    —C(═O)R⁹, -alkylR⁹, C(═O)A, C(═O)NH₂, C(═O)NHA, C(═O)NA₂, or    —C(═O)OA.

The term “hydrocarbyl” refers to any structure comprising only carbonand hydrogen atoms up to 14 carbon atoms.

The term “alkyl” used alone or as a suffix or prefix, refers to straightor branched chain hydrocarbyl radicals comprising 1 to about 12 carbonatoms.

The term “alkenyl” refers to straight or branched chain hydrocarbylradicals having at least one carbon-carbon double bond and comprising atleast 2 up to about 12 carbon atoms.

The term “alkynyl” refers to straight or branched chain hydrocarbylradicals having at least one carbon-carbon triple bond and comprising atleast 2 up to about 12 carbon atoms.

The term “cycloalkyl” refers to ring-containing hydrocarbyl radicalscomprising at least 3 up to about 12 carbon atoms.

The term “cycloalkenyl” refers to ring-containing hydrocarbyl radicalshaving at least one carbon-carbon double bond and comprising at least 3up to about 12 carbon atoms.

The term “cycloalkynyl” refers to ring-containing hydrocarbyl radicalshaving at least one carbon-carbon triple bond and comprising about 7 upto about 12 carbon atoms.

The term “aromatic” refers to hydrocarbyl radicals having one or morepolyunsaturated carbon rings having aromatic character, (e.g., 4n+2delocalized electrons) and comprising 6 up to about 14 carbon atoms.

The term “aryl” refers to aromatic radicals including both monocyclicaromatic radicals comprising 6 carbon atoms and polycyclic aromaticradicals comprising up to about 14 carbon atoms.

The term “alkylene” refers to divalent alkyl moieties, wherein saidmoiety serves to link two structures together.

The term “heterocycle” or “heterocyclic” or “heterocyclic moiety” refersto ring-containing monovalent and divalent radicals having one or moreheteroatoms, independently selected from N, O and S, as part of the ringstructure and comprising at least 3 and up to about 20 atoms in therings. Heterocyclic moieties may be saturated or unsaturated, containingone or more double bonds, and heterocyclic moieties may contain morethan one ring.

The term “heteroaryl” refers to heterocyclic monovalent and divalentradicals having aromatic character.

Heterocyclic moieties include for example monocyclic moieties such as:aziridine, oxirane, thiirane, azetidine, oxetane, thietane, pyrrolidine,pyrroline, imidazolidine, pyrazolidine, dioxolane, sulfolane2,3-dihydrofuran, 2,5-dihydrofuran tetrahydrofuran, thiophane,piperidine, 1,2,3,6-tetrahydro-pyridine, piperazine, morpholine,thiomorpholine, pyran, thiopyran, 2,3-dihydropyran, tetrahydropyran,1,4-dihydropyridine, 1,4-dioxane, 1,3-dioxane, dioxane, homopiperidine,2,3,4,7-tetrahydro-1H-azepine homopiperazine, 1,3-dioxepane,4,7-dihydro-1,3-dioxepin, and hexamethylene oxide. In additionheterocyclic moieties include heteroaryl rings such as: pyridyl,pyrazinyl, pyrimidinyl, pyridazinyl, thienyl, furyl, pyrrolyl,imidazolyl, thiazolyl, oxazolyl, pyrazolyl, isothiazolyl, isoxazolyl,1,2,3-triazolyl, tetrazolyl, 1,2,3-thiadiazolyl, 1,2,3-oxadiazolyl,1,2,4-triazolyl, 1,2,4-thiadiazolyl, 1,2,4-oxadiazolyl, 1,3,4-triazolyl,1,3,4-thiadiazolyl, and 1,3,4 oxadiazolyl. Additionally, heterocyclicmoieties encompass polycyclic moieties such as: indole, indoline,quinoline, tetrahydroquinoline, isoquinoline, tetrahydroisoquinoline,1,4-benzodioxan, coumarin, dihydrocoumarin, benzofuran,2,3-dihydrobenzofuran, 1,2-benzisoxazole, benzothiophene, benzoxazole,benzthiazole, benzimidazole, benztriazole, thioxanthine, carbazole,carboline, acridine, pyrolizidine, and quinolizidine.

In addition to the polycyclic heterocycles described above, heterocyclicmoieties include polycyclic heterocyclic moieties wherein the ringfusion between two or more rings comprises more than one bond common toboth rings and more than two atoms common to both rings. Examples ofsuch bridged heterocycles include quinuclidine,diazabicyclo[2.2.1]heptane and 7-oxabicyclo[2.2.1]heptane.

The term “halo” or “halogen” refers to fluorine, chlorine, bromine andiodine radicals.

The term “alkoxy” refers to radicals of the general formula —O—R,wherein R is selected from a hydrocarbyl radical. Alkoxy moietiesinclude methoxy, ethoxy, propoxy, isopropoxy, butoxy, t-butoxy,isobutoxy, cyclopropylmethoxy, allyloxy, and propargyloxy.

The term amine or amino refers to radicals of the general formula —NRR′,wherein R and R′ are independently selected from hydrogen or ahydrocarby radical.

DETAILED DESCRIPTION OF THE INVENTION

In a further aspect of the invention, A, R¹ and R³, each independently,as an alkyl, alkenyl, alkynyl and as a cycloalkyl, may optionally besubstituted with halogen, nitro, cyano, hydroxy, trifluoromethyl, amino,carboxy, carboxamido, amidino, carbamoyl, mercapto, sulfamoyl, C₁₋₄alkyl, C₂₋₄ alkenyl, C₂₋₄ alkynyl, C₃₋₆ cycloalkyl, C₃₋₆ cycloalkenyl,C₁₋₄ alkoxy, C₁₋₄ alkanoyl, C₁₋₄ alkanoyloxy, N-(C₁₋₄ alkyl), N(C₁₋₄alkyl)₂, C₁₋₄ alkanoylamino, (C₁₋₄ alkanoyl)₂amino, N—(C₁₋₄alkyl)carbamoyl, N,N—(C₁₋₄ alkyl)₂carbamoyl, (C₁₋₄)S, (C₁₋₄ alkyl)S(O),(C₁₋₄alkyl)S(O)₂, (C₁₋₄) alkoxycarbonyl, N—(C₁₋₄ alkyl)sulfamoyl,N,N—C₁₋₄ alkyl)sulfamoyl, C₁₋₄ alkylsolfonylamino, and heterocyclic.

A, R¹ and R³ each independently as an alkyl, alkenyl or alkynyl may bestraight or branched, preferably having 1-6 carbon atoms. A, R¹ and R³preferably have 3-6 atoms when each are independently a cyclic alkyl.Other preferable values for A, R¹ and R³ when each are an alkyl includemethyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl,cyclopentyl, neopentyl and cyclohexyl. Preferable values for R¹ when R¹is a halogen are fluorine, chlorine, and bromine. Other preferablevalues for R¹ when R¹ is at position 6 on the bicyclic ring are methyl,ethyl, ethoxy and methoxy. Preferable values for R¹ when R¹ is atposition 5 on the bicyclic ring are —H, methyl, ethyl and methoxy. WhenR¹ is at position 5- on the bicyclic ring, R¹ is more preferably —H.When R¹ is at position 7- on the bicyclic ring, R¹ is preferably —H.

Particular values for R² are substituent groups of Formula i. PreferablyR² is represented by formula i, wherein n equals 2. Most preferably R²is represented by N-methyl piperazinyl.

Particular values for R³ are hydrogen, methyl, ethyl, n-propyl,isopropyl, n-butyl, isobutyl, and tert-butyl. R³ is preferably methyl.

Particular values for R⁴ are hydrogen, methyl, ethyl, n-propyl,isopropyl and trimethylsilanyl-ethoxymethoxy. Preferably, R⁴ is methyl.

R⁶ is preferably represented by H.

Y represents a linking group. Y is preferably —C(═O)N(CH₃)—, when Y is—C(═O)N(A)-. Y may also be —C(═O)-piperazine. When Y represents afive-membered heterocyclic ring, Y may be represented by, for example,pyrrole, thiophene, furan, imidazole, thiazole, oxazole, pyrazole,isothiazole, isoxazole, 1,2,3-triazole, 1,2,3-thiadiazole,1,2,3-oxadiazole, 1,2,4-triazole, 1,2,4-thiadiazole, 1,2,4-oxadiazole,1,3,4-triazole, 1,3,4-thiadiazole or 1,3,4-oxadiazole.

More preferably, Y is —C(═O)NH—.

Examples of R⁷ that represent monocyclic or bicyclic aromatic ring or aheterocycle include, but are not limited to, phenyl; 1- and 2-naphthyl;2-, 3- and 4-pyridyl; 2- and 3-thienyl; 2- and 3-furyl1-, 2- and3-pyrrolyl, imidazolyl, thiazolyl, oxazolyl, pyrazolyl, isothiazolyl,isoxazolyl, 1,2,3-triazolyl, 1,2,3-thiadiazolyl, 1,2,3-oxadiazolyl,1,2,4-triazolyl, 1,2,4-thiadiazolyl, 1,2,4-oxadiazolyl, 1,3,4-triazolyl,1,3,4-thiadiazolyl, 1,3,4 oxadiazolyl, quinolyl; isoquinolyl; indolyl;benzothienyl and benzofuryl, benzimidazolyl, benzthiazolyl,benzoxazolyl, or triazinyl.

R⁷ may also be represented by the Formula (v):

R⁷ may further be represented by the Formula (vi):

When the values for R⁷ are as set forth above, R⁸ may be a single bondas tether, —C(═O)—, —CH₂—, —C(═O)—, —SO₂—, —S(═O)—, —S—, —O—, —C(═O)NH—,—SO₂NH—, or a five membered heterocycle connected to R⁷ by a ringfusion; and R⁹ may represent an aryl, heterocyclic or heteroaryl eachindependently optionally substituted with halogen, nitro, cyano,hydroxy, trifluoromethyl, amino, carboxy, carbamoyl, mercapto,sulfamoyl, C₁₋₄ alkyl, C₂₋₄alkenyl, C₂₋₄ alkynyl, C₃₋₆ cycloalkyl, C₃₋₆cycloalkenyl, C₁₋₄ alkoxy, C₁₋₄ alkanoyl, C₁₋₄ alkanoyloxy,N—(C₁₋₄alkyl), N(C₁₋₄ alkyl)₂, C₁₋₄ alkanoylamino, (C₁₋₄alkanoyl)₂amino, N—(C₁₋₄alkyl)carbamoyl, N,N-(C₁₋₄)₂carbamoyl, C₁₋₄)S,C₁₋₄S(O), (C₁₋₄alkyl)S(═O)₂, (C₁₋₄) alkoxycarbonyl, N—(C₁₋₄alkyl)sulfamoyl, N,N—C₁₋₄ alkyl)sulfamoyl, C₁₋₄ alkylsolfonylamino, orheterocyclic. Preferably R⁹ is a heterocyclic moiety.

More preferably R⁹ represents piperazine, thiomorpholine or morpholineoptionally substituted on carbon with at least one substituent selectedfrom A. R⁸ may be a five membered heterocycle, incorporating at leastone heteroatom selected from N, O, or S and it may be connected to R⁷ bya ring fusion, preferably when R⁷ is phenyl. When R⁸ is a single bond astether, R⁹ is preferably methoxy, cyano, a five-membered heterocycle ora compound represented by the Formula (vii):

When R⁸ is represented by a 5-membered heterocyclic comprising N andfurther when it is connected to R⁷ by a ring fusion, R⁹ is preferably—C(═O)A attached at the nitrogen atom. R⁹ is most preferably—C(═O)CH₂CH₃.

When R⁷ is phenyl or a 6-membered heterocyclic ring, R⁹, is attached viathe R⁸ tether at the 2-, 3- or 4-position of the phenyl or a 6-memberedheterocyclic ring. Preferably, R⁹ is attached via the R⁸ tether at the3- or 4-position of the phenyl or a 6-membered heterocyclic ring. Morepreferably, R⁹ is attached via the R⁸ tether at the 4 position of thephenyl or a 6-membered heterocyclic ring.

R¹⁰ may be represented by alkyl or cycloalkyl each independentlyoptionally substituted with halogen, nitro, cyano, hydroxy,trifluoromethyl, amino, carboxy, carbamoyl, mercapto, sulfamoyl, C₁₋₄alkyl, C₂₋₄alkenyl, C₂₋₄ alkynyl, C₃₋₆ cycloalkyl, C₃₋₆ cycloalkenyl,C₁₋₄ alkoxy, C₁₋₄ alkanoyl, C₁₋₄ alkanoyloxy, N—(C₁₋₄ alkyl), N(C₁₋₄alkyl)₂, C₁₋₄ alkanoylamino, (C₁₋₄ alkanoyl)₂amino,N—(C₁₋₄alkyl)carbamoyl, N,N—(C₁₋₄)₂carbamoyl, C₁₋₄)S, C₁₋₄S(O),(C₁₋₄alkyl)S(O)₂, (C₁₋₄) alkoxycarbonyl, N—(C₁₋₄ alkyl)sulfamoyl,N,N—C₁₋₄ alkyl)sulfamoyl, C₁₋₄ alkylsulfonylamino, or heterocyclic. R¹⁰is preferably a halogen, preferably chlorine or fluorine, cyano, or—OCH₃. When R¹⁰ is a halogen it is preferably chlorine or fluorine. WhenR⁷ is a phenyl or 6-membered heteroaromatic ring, R¹⁰ is attached at the2-, 3- or 4-position of the phenyl or a 6-membered heterocyclic ring.Preferably, R¹⁰ is attached at the 2- or 3-position of the phenyl or a6-membered heterocyclic ring when R⁹ is attached via the R⁸ tether atthe 4-position of the phenyl or a 6-membered heterocyclic ring. Morepreferably, R¹⁰ is attached at the 3-position of the phenyl or a6-membered heterocyclic ring when R⁹ is attached via the R⁸ tether atthe 4-position of the phenyl or a 6-membered heterocyclic ring.

When R⁸ is represented by a single bond as tether, R⁹ is preferablyrepresented by piperazine, thiomorpholine or morpholine optionallysubstituted on carbon with at least one substituent selected from A.

Also provided herein is when R⁸ is a single bond tether and R⁹ ispiperzine-R¹¹. When R¹¹ represents SO₂A it is preferably represented byan alkylsufonyl, more preferably —SO₂CH₃, —SO₂CH₂CH₃, SO₂-n-C₃H₇,SO₂-i-C₃H₇, SO₂-n-C₄H₁₀, —SO₂-i-C₄H₁₀, or —SO₂-t-C₄H₁₀. When R¹¹represent C(═O)A, it is preferably represented by an alkylcarbonyl morepreferably —(═O)CH₃, —C(═O)CH₂CH₃, C(═O)-n-C₄H₁₀, —C(═O)-i-C₄H₁₀,—C(═O)-t-C₄H₁₀, or —C(═O)C₃H₇. — When R¹¹ is represented by C(═O)NHA orC(═O)NA₂ it is preferably an alkyl or dialkyl carbamoyl more preferablyC(═O)NCH₂CH₃, C(═O)NH-cycloC₆H₁₂, or C(═O)NH-cycloC₅H₁₀. When R¹¹ isrepresented by C(═O)R⁹ it is preferably —C(═O)-pyrrolidine, or—C(═O)-morpholine. When R¹¹ is represented by SO₂NA₂ it is preferablySO₂N(CH₃)₂. When R¹¹ is represented by AOH, it is preferably representedby, CH₂CH₂OH or —(═O)CH₂CH₂OH. R¹¹ may also be represented by—C(═O)OC₄H₁₀.

The compounds provided herein are useful in the form as a free base, butmay also be provided in the form of a pharmaceutically acceptable salt,and/or in the form of a pharmaceutically acceptable hydrate. For examplepharmaceutically acceptable salts of compounds of Formula I includethose derived from mineral acids such as for example: hydrochloric acid,nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid,hydroiodic acid, nitrous acid, and phosphorous acid. Pharmaceuticallyacceptable salts may also be developed with organic acids includingaliphatic mono and dicarboxylates and aromatic acids. Otherpharmaceutically-acceptable salts of compounds of the present inventioninclude for example hydrochloride, sulfate, pyrosulfate, bisulfate,bisulfite, nitrate, and phosphate.

Processes for the manufacture of the compounds of Formula I are providedas further features of the invention. Many of the Compounds describedherein can be made by processes known in the chemical arts for theproduction of structurally analogous compounds. Accordingly, thecompounds of this invention may be prepared by employing proceduresknown in the literature starting from known compounds or readilyprepared intermediates.

For compounds of the present invention that have Y as an amide linker,the compounds are preferably made by the general procedure for amidecoupling, that is by coupling an anime with an acid hydrochloride. Theamines used in the current invention if not commercially available maybe made by known techniques. For example as a first step in the processof making compound of Formula I, a nitro compound may be reduced to anamine. The nitro compound may be a nitrophenyl compound. The resultingamines may be reacted with an acid hydrochloride. A method for preparingthe acid hydrochlorides useful in synthesis of chromones is set forth inScheme 1 on the following page.

Alternatively, the chromone-2-carboxylic acid may be converted to theacid chloride and reacted immediately with an appropriate amine, asdepicted in Scheme 2, below. Additional functional group manipulationsinclude, but are not limited to, O-dealkylation and N-dealkylation(Scheme 3).

Quinoline and quinolone compounds of the present invention are preparedand derivatized via synthetic routes similar to those employed forsynthesis of the chromone-2-carboxamides described above and in Schemes1-3. These synthetic routes to quinoline and quinolone compounds of thepresent invention are depicted in Scheme 4, infra.

invention contain for example asymmetrically substituted carbon and/orsulfur atoms, and accordingly may exist in and be isolated in,optically-active and racemic forms. Some compounds may exhibitpolymorphism, thus it is to be understood that the present inventionencompasses racemic, optically-active, polymorphic or stereoisomericforms, or mixtures thereof, which forms possess properties useful in thetreatment of the disorders set forth below. Preparation of opticallyactive forms is well known in the art how (for example by resolution ofracemic forms by recrystallization techniques, synthesis fromoptically-active starting materials, chiral synthesis, or bychromatographic separation using a chiral stationary phase) and how todetermine efficacy for the treatment of the disorder described above.

Compounds of Formula I have been found by the inventors to be useful as5-HT_(1B) and 5HT_(1D) antagonists. The compounds of Formula I, andtheir pharmaceutically acceptable salts, may also be used in a methodfor the treatment of depression, generalized anxiety, eating disorders,dementia, panic disorder, sleep disorders, gastrointestinal disorders,motor disorders, endocrine disorders, vasospasm and sexual dysfunction.The treatment of these disorders comprises administering to awarm-blooded animal, preferably a mammal, more preferably a human, inneed of such treatment, an effective amount of a compound of Formula I,or a pharmaceutically acceptable salt of said compound.

Further provided herein are compounds of Formula I, and theirpharmaceutically acceptable salts, for use in the treatment ofdepression, generalized anxiety, eating disorders, dementia, panicdisorder, sleep disorders, gastrointestinal disorders, motor disorders,endocrine disorders, vasospasm and sexual dysfunction of a warm-bloodedanimal, preferably a mammal, more preferably a human, in need of suchtherapy.

Further provided herein is a method of treatment of a warm-bloodedanimal, preferably a mammal, more preferably a human, suffering fromdisorders such as depression, generalized anxiety, eating disorders,dementia, panic disorder, sleep disorders, gastrointestinal disorders,motor disorders, endocrine disorders, vasospasm and sexual dysfunctioncomprising administering to such animal an effective amount of acompound of Formula I or, or a pharmaceutically acceptable salt of thecompound.

Further provided is the use of a compound of Formula I in thepreparation of a medicament for the treatment of a disorder such asdepression, generalized anxiety, eating disorders, dementia, panicdisorder, sleep disorders, gastrointestinal disorders, motor disorders,endocrine disorders, vasospasm and sexual dysfunction in a warm-bloodedanimal, preferably a mammal, more preferably a human, suffering fromsuch disorder.

The invention further provides a pharmaceutical composition suitable forthe treatment of the above describe disorders comprising administeringto a warm-blooded animal having such disorder an effective amount of apharmaceutical composition of a compound of Formula I, or apharmaceutically acceptable salt.

The invention also provides a pharmaceutical composition comprising acompound of Formula I as defined herein, or a pharmaceuticallyacceptable salt, in combination with a pharmaceutically acceptablecarrier. Preferred compounds of Formula I for use in the compositions ofthe invention are as described above.

All compounds described herein demonstrate binding affinities (observedKi values), in an assay described below, of less than about 10 μM.Further, compounds of the present invention not only demonstrate5HT_(1B) antagonist activity by reversing 5HT_(1B) agonist-inducedhypothermia in the guinea pig, these compounds are considered to beorally active, and hence, they are the preferred compounds. Examples 1,10, 11, 31, 32, 34, 44, 55, 56, 57, 71 and 72, infra, demontrate5HT_(1B) antagonist activity in a dosage range of 0.006-5.5 mg/kg. Inaddition, compounds described herein demonstrate activity in the learnedhelplessness assay for antidepressant/antianxiety activity. Examples 31,44, 71 and 72, infra, demonstrate activity in the learned helplessnessassay. In addition, compounds were tested for maximal intrinsic activity(IA), and were found to have measured IA's of negative 50% to positive150% in the GTPγS assay described below, thus demonstrating a range ofresponse from agonism (low percentages) to antagonism (highpercentages).

The compounds described herein may be provided or delivered in a formsuitable for oral use, for example in a tablet, lozenge, hard and softcapsule, aqueous solution, oily solution, emulsion, and suspension. Thecompounds may be also be provided for topical administration, forexample, as a cream, ointment, gel, spray, or aqueous solutions, oilysolutions, emulsions or suspensions. The compounds described herein mayalso be provided in a form suitable for nasal administration forexample, as a nasal spray, nasal drops, or dry powder. The compositionsmay also be administered to the vagina or rectum in the form of asuppository. The compounds described herein may also be administeredparentally, for example by intravenous, intravesicular, subcutaneous, orintramuscular injection or infusion. The compounds may be administeredby insufflation (for example as a finely divided powder). The compoundsmay also be administered transdermally or sublingually.

The compositions of the invention may accordingly be obtained byconventional procedures using conventional pharmaceutical excipients,well known in the art. Thus, compositions intended for oral use maycontain, for example, one or more coloring, sweetening, flavoring and/orpreservative agents.

The amount of active ingredient that is combined with one or moreexcipients to produce a single dosage form will necessarily varydepending upon the host treated and the particular route ofadministration. The size of the dose for therapeutic or prophylacticpurposes of a compound of the Formula I will naturally vary according tothe nature and severity of the conditions, the age and sex of the animalor patient and the route of administration, according to well knownprinciples of medicine. Various assays and in vivo tests are known fordetermining the utility of the compounds in the disorders noted aboveand specifically as agonists and antagonists of 5HT_(1B) and 5H_(1D).

The utility of the compounds for example to treat depression may beshown via a learned helplessness test in guinea pigs, which is usedextensively as correlative to antidepressant activity in humans. Thelearned helplessness test may be carried out as follows: Seventy maleHartley guinea pigs, each weighing about 350-425 gm are fed ad lib, andare housed under a 12-hour light/dark cycle. The procedure consists oftwo phases: The induction phase and the avoidance-training phase. In theinduction phase, subjects are placed into standard shuttle cages (20L×16 W×21 centimeters H) which are fitted with a grid floor. Electricalstimulation (1.25 mA, 10 sec duration) is delivered to the floor of thecage every 90-sec during 1 hour daily sessions. Subjects have noopportunity to escape or to avoid shocks. Induction is conducted for 2consecutive days.

In avoidance training, testing is also conducted in the shuttle cages,except that the subjects are not returned to the same chamber in whichinduction had occurred. Additionally, all cages are fitted with apartition with an arch in the center of the cage, through which animalscan pass between the left and right halves of the cage. The procedureemployed is a standard shuttle avoidance procedure in which a compound,conditioned stimulus (a 10-sec presentation of a tone and turning on ofa lamp on the side of the cage that the guinea pig was occupying) servesto indicate presentation of electrical current to the floor of the cage.Shock is presented for a 5 sec period, 5 sec after initiation of theconditioned stimulus. Entry into the opposite side of the shuttle cagevia the arched partition prior to shock onset results in the end of thetrial (avoidance response). If shock is delivered, entry into theopposite side of the cage results in termination of the shock and CS(escape). Reversal of learned helplessness in the induction subjectscorrelates to antidepressant activity of the test compound.

Avoidance training, 45-min in duration, is conducted on 2 consecutivedays, beginning 48 hr after the final induction session. Seventysubjects are assigned to 1 of 6 groups of 11-12 animals. The groups areas follows:

-   -   1) No induction group. The subjects are placed into the shuttle        cages but are not given inescapable shock, the animals are        subsequently trained in the avoidance procedure and the vehicle        is administered;    -   2) Induction vehicle control group;    -   3) Imipramine 17.8 mg/kg;    -   4) 0.3 mg/kg compounds;    -   5) 1 mg/kg compounds; and    -   6) 5 mg/kg compounds.

Groups 2-6 are given induction and avoidance training sessions.Injections are administered immediately following induction sessions and1 hour prior to avoidance training sessions. A second injection isadministered 7-8 hours following the first injection, for a total of 9injections administered over 5 days. No injections are administeredfollowing the final avoidance training session.

Compounds of the present invention may be administered in a volume of 1mL/kg bwt. Imipramine is dissolved in DI water. The compounds aredissolved in DI water, to which was added a few drops of lactic acid (pH5.5). The vehicle control is DI water prepared with lactic acid to thesame pH as the treated groups.

The primary dependent variable is escape failure during avoidancetraining. 2-way analysis of variance (ANOVA) is used to assess overalltreatment effect, with Dunn's post hoc analysis used to compare thevehicle-treated group with the drug-treated groups. The no-inductiongroup is used to gauge whether learned helplessness is established, bycomparison to the vehicle treated group.

An alternative method for determining the utility of the compounds ofthe present invention is to investigate the in vivo activity of thecompounds using a guinea pig hypothermia test (J. Med. Chem., 41:1218-1235 (1998)). Compounds that bind to 5-HT_(1B) receptors are knownto be useful in treating disorders described above (e.g., depression,generalized anxiety, eating disorders, dementia, panic disorder, sleepdisorders, gastrointestinal disorders, motor disorders, endocrinedisorders, vasospasm and sexual dysfunction. While not wishing to bebound to any theory, it is believed that 5-HT_(1B) receptors on nerveterminals control the amount of release of s5-ht into the synapse. Thus,it can be shown that compounds of Formula I, and their pharmaceuticallyacceptable salts, are able to act as 5-HT_(1B) antagonists and block theagonist-induced effect of hypothermia (a drop in body temperature ofabout 2° C. observed within 0.5-1.5 hours following administration of a5-HT_(1B) agonist) as a method for assessing whether the novel compoundsare effective as antagonists at the 5-HT_(1B) receptor.

The hypothermia test is conducted as follows: A tele-thermometer fittedwith a flexible probe will be used. The tip of the probe is immersed ina test tube containing a lubrication agent between usage. Coretemperature is measured by inserting the probe into the rectum and bywaiting for the temperature to stabilize, which occurs within the 20-60seconds. Core temperature is measured once (pretest) prior toadministration of the test substance in order to establish a baselinetemperature for all animals. Guinea pigs are then dosed with the testsubstance (candidate 5-ht1b antagonist) either subcutaneously orintraperitoneally. In general, 30 min following dosing with antagonist,agonist is administered subcutaneously. The temperature is then recorded30-, 60, 90-min following agonist. In some studies, in order to recordtime course of antagonist activity, up to 12 hours may be allowed toelapse between administration of antagonist and agonist. The drugs mayeither be injected subcutaneously, intraperitoneally or orally (using aflexible plastic gavage tube, or a stainless steel gavage tube). Inaddition, animals may be observed on the days following drugadministration in order to monitor for unexpected toxicity. The bodytemperature of the guinea pigs is recorded separately for each guineapig at each test time point, and submitted to a ANOVA with one betweensubjects factor: dose, and one within subject factor: time. Following asignificant two-way interaction (p<0.05), Dunnett's t-test is performedto compare the drug treatment with either the saline or the effects oftreatment with the hypothermic agent.

Male Guinea Pig (Dunkin-Hartley), maximum 3 animals per cage, are used.The animals may be grouped in sets of 5 during testing. The animals willnot be deprived of food or water during their time in the laboratory.The routes of administration are: S.C., I.P., P.O. The maximum dose(volume) is 2 ml/kg s.c. or i.p., 5 ml/kg P.O. three times daily.

This method may function as a primary in vivo screen for compoundshaving an affinity for 5-ht_(1b) receptors as a determination ofantagonist activity. Each experiment may consist of separate groups of 5subjects per treatment level. One group is given vehicle prior toagonist administration and may serve as the control group, i.e.,hypothermia will be unaltered by introduction of an antagonist. Theother groups are administered different doses of antagonist prior toagonist administration, but no more than 5 groups are tested at a time.In order to determine full dose effect functions for compounds (todetermine drug potency) 4-6 doses of each compound are evaluated. Thatresults in about 25-35 animals per drug to be evaluated. Dose-responsecurves are generated and ED50 values are determined. ED50 values forcompounds of the present invention range from 0.006-5.5 mg/kg.

Other assays that may be used to measure for example affinity ofcompounds of the present invention for 5HT_(1B) and 5HT_(1D) receptorsare described in J. Med. Chem 41:1218-1235, 1228 (1998) and J. Med. Chem42:4981-5001, (1999) and incorporated by reference herein. These assaysmay be used with some modifications: Frozen membrane preparations of astably transfected chinese hamster ovary (CHO) cell line expressing5-HT_(1B) receptors and 5-HT_(1D) receptors are thawed rapidly, brieflyvortexed, and diluted in assay buffer (AB) containing 50 mM Tris-HCl, 4mM MgCl₂, 4 mM CaCl₂, 1 mM EDTA, and adjusted to pH 7.4 with NaOH. Finalprotein concentrations are −0.185 mg/ml for 5-HT_(1B), and 0.4 mg/ml for5-HT_(1D) membranes. Test compounds are evaluated in competition assaysusing [³H]-GR125743 (Amersham). The ligand concentration in both assayswas 0.27 nM. Kd for [³H]-GR125743 may vary from 0.15 nM to 0.25 nM. The5-HT_(1B) and 5-HT_(1D) assays are performed simultaneously on one96-well assay plate, one drug/compound per plate. Ten serial dilutions(1 uM to 4 pM, final concentration) of compound are prepared in DMSOfrom 10 mM stock solutions. Incubation mixtures are prepared inquadruplicate in 96-deep well assay plates (Matrix 1 ml). Final assayvolumes per well are 10 μl compound/nonspecific; 100 μl membranes; 100μl [3H]-GR125743; and 790 μl AB. Specific binding is defined by using 10uM Methiothepine. The assay plates are shaken for 5 min., and thenincubated for an additional 55 min. Then the assay plates are filteredthrough Beckman GF/B filters (soaked>2 hrs. in PEI) using a PackardFiltermate 196. Filters are washed 2× with 1 ml ice-cold wash buffer (5mM Tris-HCl-pH7.4 with NaOH). After the filters are dried, 35 μl ofMicroscint20 is added to each well. The plates are then counted on aPackard TopCount to determine CPM's per well. Ki values are determinedfor each test compound utilizing the graphic and analytical softwarepackage, GraphPad Prism. Compounds are then ranked in order of potency,and selectivity for 5-HT_(1B) over 5-HT_(1D) receptors.

A method that may be used to determine a compound's affinity for5-HT_(1B) and 5HT_(1D) receptors is a guinea pig cortical test. Thisassay is described in detail by Roberts, et al, Br. J. Pharmacol., 1996,117, 384-388, which is incorporated by reference herein. The test iscarried out as follows: Guinea pigs are decapitated and the cortici isdissected out, weighed and homogenized in 50 mM Tris-HCl, pH 7.7 with anUltra-Turrax followed by centrifugation for 10 min at 48000×g and 5° C.The pellet is resuspended and recentrifuged. The final pellet issuspended in 0.32 M sucrose buffer to a concentration of 0.5 g originalwet weight per mL and stored frozen at −70° C. The radioligand bindingassay is carried out as follows: [³H]GR125743 saturation studies aretested in duplicate with 3-4 mg w.w. per tube in 5 mL buffer (50 mMTris, 4 mM CaCl2, 4 mM MgCl2 and 1 mM EDTA at pH 7.7), and aconcentration range of 0.012-2 nM (10-12 concentrations) for theradioligand. Non-specific binding is determined in the presence of 10 mMmethiothepin. In competition experiments 4-8 mg w.w. per tube and aradioligand concentration of 0.2 nM are used with 10-12 concentrationsof the competing drug. The assays are run for ₂₋₄ hours at 30° C. andterminated by rapid filtration through Whatman GF/B filters (pretreatedwith 0.1% polyethyleneimine) using a Brandel cell harvester. Bovineserum albumin (0.1%) is added to the washing buffer to reducenon-specific binding. Data from the experiments may be analyzed usingthe iterative non-linear curve-fitting program LIGAND. The K_(d) valuesobtained from the saturation studies are used in the calculation of theKi values by the LIGAND program. The K_(d) value of [³H]GR125743 mayresult in a measurement of 46±4 pM and the B_(max) in a measurement of4.9±0.2 pmol/g w.w.

A GTPγS binding assay may used to determine whether a compound is a5HT_(1B) or 5HT_(1D) agonist or antagonist. One assay available measuresagonist stimulated GTP binding for example as set forth by Lazareno, S.(1999) Methods in Molecular Biology 106: 231-245. Membrane preparationsof a stably transfected CHO cell line expressing human 5-HT_(1B)receptors are purchased for example from Unisyn, Hopkinton, Mass. Frozenmembranes are thawed, briefly sonicated, and diluted to 167 μg/mlprotein in assay buffer containing 20 mM HEPES, 100 mM NaCl, 1 mM MgCL₂and 1 μM GDP, pH adjusted to 7.4 with NaOH. Diluted membranes arebriefly homogenized with a Polytron and allowed to equilibrate at roomtemperature for at least 15 minutes before use. Serial dilutions (10 μMto 1 pM, final concentration) of test compounds are prepared in bufferwith and without 100 nM 5-HT (final concentration) from 10 mM DMSO stocksolutions. Incubation mixtures are prepared in quadruplicate in 96-well,deep-well plates and consisted of 180 μL of membranes (30 μg protein)and 40 μL of compound with or without 5-HT. After an incubation periodof 15 minutes at room temperature, 20 μL of [³⁵S]GTPγS (NEN; 100 pMfinal concentration) is added to begin the assay. Mixtures are shakenfor 2 minutes and incubated at room temperature for an additional 28minutes. The reaction is stopped by rapid filtration through BeckmanGF/B glass fiber filters using a 96-well Packard cell harvester. Filtersare washed four times with 1 mL ice-cold water. The filter plates arenominally dried and 30 μL of scintillation cocktail (MicroScint 40,Packard) is added to each well. CPMs for each well is determined using aTopCount Scintillation Counter (Packard). Maximum stimulation of[³⁵S]GTPγS binding is defined in the presence of 100 nM 5-HT. Basal[³⁵S]GTPγS binding is defined in buffer alone. IC50 values are definedas the concentration of compound at which 50% of the 100 nM 5-HTresponse [was] obtained. Maximal intrinsic activity (IA) of a compoundis defined as the percent maximal 5-HT-induced stimulation by 10 μMcompound in the absence of 5-HT. As an inter-assay standard, aconcentration response curve of 5-HT (1 μM to 1 pM final) in the absenceof compounds was included in each assay and an EC₅₀ was determined.

Preferred compounds of the present invention include, but are notlimited to, the following compositions listed in Table 1 on thefollowing pages. TABLE 1 Compounds of the present invention Example #Structure Name 1

8-(4-methyl-1-piperazinyl)-N-[4-(4-morpholinyl)phenyl]-4-oxo-4H-chromene-2- carboxamide 2

2-{1-[4-(2-Methoxy-phenyl)-piperazin-1-yl]- methanoyl}-8-(4-methyl-piperazin-1-yl)- chromen-4-one 3

2-{1-[4-(1-Acetyl-2,3-dihydro-1H-indol-6-yl)-piperazin-1-yl]-methanoyl}-8-(4-methyl- piperazin-1-yl)-chromen-4-one 4

2-Chloro-5-(4-{1-[8-(4-methyl-piperazin-1-yl)-4-oxo-4H-cbromen-2-yl]-methanoyl)- piperazin-1-yl)-benzonitrile 5

2-{1-[4-(4-Methoxy-phenyl)-piperazin-1-yl]-methanoyl}-8-(4-methyl-piperazin-1-yl)- chromen-4-one 6

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H- chromene-2-carboxylic acid(5-furan-2-yl- 1H-pyrazol-3-yl)-amide 7

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H- chromene-2-carboxylic acid(4-imidazol- 1-yl-phenyl)-amide 8

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H- chromene-2-carboxylic acid (4-[1,2,3]thiadiazol-5-yl-phenyl)-amide 9

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H- chromene-2-carboxylic acid 4-[1,2,3]thiadiazol-5-yl-benzylamide 10

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H- chromene-2-carboxylic acid[4-(4-acetyl- piperazin-1-yl)-phenyl]-amide 11

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H- chromene-2-carboxylic acid [4-(4-methanesulfonyl-piperazin-1-yl)-phenyl]- amide 12

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H- chromene-2-carboxylic acid(2-methoxy-4- morpholin-4-yl-phenyl)-amide 13

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H- chromene-2-carboxylic acid(3-chloro-4- morpholin-4-yl-phenyl)-amide 14

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H- chromene-2-carboxylic acid(4-thiomorpholin- 4-yl-phenyl)-amide 15

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H- chromene-2-carboxylic acid(2,5-diethoxy-4- morpholin-4-yl-phenyl)-amide 16

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H- chromene-2-carboxylic acid(4-cyanomethyl- phenyl)-amide 17

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H- chromene-2-carboxylic acid(1H-indol-5-yl)- amide 18

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H- chromene-2-carboxylic acid[4-(1-morpholin- 4-yl-methanoyl)-phenyl]-amide 19

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H- chromene-2-carboxylic acid[4-(2,6- dimethyl-morpholin-4-yl)-phenyl]-amide 20

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H- chromene 2-carboxylic acid[4-(4-fluoro- phenoxy)-phenyl]-amide 21

8-(4-Methyl-piperazin-1-yl)-2-(6- morpholin-4-yl-benzooxazol-2-yl)-chromen-4-one 22

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H- chromene-2-carboxylic acid(2-hydroxy-4- morpholin-4-yl-phenyl)-amide 23

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H- chromene-2-carboxylic acid(5-ethoxy- benzothiazol-2-yl)-amide 24

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H- chromene-2-carboxylic acid(4-bromo- phenyl)-amide 25

8-(4-Methylpiperazin-1-yl)-4-oxo-4H- chromene-2-carboxylic acid methyl-(4-morpholin-4-yl-phenyl)amide 26

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H- chromene-2-carboxylic acid (3-morphohn-4-yl-phenyl)-amide 27

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H- chromene-2-carboxylic acid (3-cyano-4-morpholin-4-yl- phenyl)-amide 28

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H- chromene-2-carboxylic acid (3-fluoro-4-morpholin-4- yl-phenyl)-amide 29

4-[4-({1-[8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromen-2-yl]-methanoyl}- amino)-phenyl]-piperazine-1- carboxylicacid tert-butyl ester 30

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H- chromene-2-carboxylic acid(4-piperazin-1-yl-phenyl)-amide 31

6-Methoxy-8-(4-methyl-piperazin-1-yl)- 4-oxo-4H-chromene-2-carboxylicacid (4-morpholin-4-yl-phenyl)-amide 32

6-Methoxy-8-(4-methyl-piperazin-1-yl)- 4-oxo-4H-chromene-2-cazboxylicacid [4-(4-methanesulfonyl-piperazin- 1-yl)-phenyl]-amide 33

6-Methoxy-8-(4-Methyl-piperazin-1-yl)- 4-oxo-4H-chromene-2-carboxylicacid (3- chloro-4-morpholin-4-yl-phenyl)-amide 34

6-Methoxy-8-(4-methyl-piperazin-1-yl)- 4-oxo-4H-chromene-2-carboxylicacid (3- fluoro-4-morpholin-4-yl-phenyl)-amide 35

6-Methoxy-8-(4-methyl-piperazin-1-yl)- 4-oxo-4H-chromene-2-carboxylicacid (2- methoxy-4-morpholin-4-yl-phenyl)- amide 36

6-Methoxy-8-(4-methyl-piperazin-1-yl)- 4-oxo-4H-chromene-2-carboxylicacid (4- thiomorpholin-4-yl-phenyl)-amide 37

6-Methoxy-8-(4-methyl-piperazin- 1-yl)-4-oxo-4H-chromene-2- carboxylicacid [4-(2,6-dimethyl- morpholin-4-yl)-phenyl]-amide 38

6-Methoxy-8-(4-methyl-piperazin- 1-yl)-4-oxo-4H-chromene-2- carboxylicacid (3-morpholin-4-yl- phenyl)-amide 39

6-Methoxy-8-(4-methyl-piperazin- 1-yl)-4-oxo-4H-chromene-2- carboxylicacid {4-[4-(2-hydroxy- ethyl)-piperazin-1-yl]-phenyl}- amide 40

6-Methoxy-8-(4-methyl-piperazin- 1-yl)-4-oxo-4H-chromene-2- carboxylicacid [4-(1-morpholin-4- yl-methanoyl)-phenyl]-amide 41

6-Methoxy-8-(4-methyl-piperazin- 1-yl)-4-oxo-4H-chromene-2- carboxylicacid (3-cyano-4- morpholin-4-yl-phenyl)-amide 42

4-[4-({1-[6-Methoxy-8-(4-methyl- piperazin-1-yl)-4-oxo-4H-chromen-2-yl]-methanoyl}-amino)-phenyl]- piperazine-1-carboxylic acid tert-butyl ester 43

6-Methoxy-8-(4-methyl-piperazin-1- yl)-4-oxo-4H-chromene-2-carboxylicacid (4-piperazin-1-yl-phenyl)-amide 44

6-Methoxy-8-(4-methyl-piperazin-1- yl)-4-oxo-4H-chromene-2-carboxylicacid [4-(4-propionyl-piperazin-1-yl)- phenyl]-amide 45

6-Methoxy-8-(4-methyl-piperazin-1- yl)-4-oxo-4H-chromene-2-carboxylicacid [4-(4-ethane sulfonyl-piperazin- 1-yl)-phenyl]-amide 46

6-Methoxy-8-(4-methyl-piperazin-1- yl)-4-oxo-4H-chromene-2-carboxylicacid [4-(4-dimethyl sulfamoyl- piperazin-1-yl)-phenyl]-amide 47

4-[4-({1-[6-Methoxy-8-(4-methyl- piperazin-1-yl)-4-oxo-4H-chromen-2-yl]-methanoyl}-amino)-phenyl]- piperazine-1-carboxylic aciddimethylamide 48

4-[4-({1-[6-Methoxy-8-(4-methyl- piperazin-1-yl)-4-oxo-4H-chromen-2-yl]-methanoyl}-amino)-phenyl]- piperazine-1-carboxylic acid ethylamide49

4-[4-({1-[6-Methoxy-8-(4-methyl- piperazin-1-yl)-4-oxo-4H-chromen-2-yl]-methanoyl}-amino)-phenyl]- piperazine-1-carboxylic acidcyclohexylamide 50

4-[4-({1-[6-Methoxy-8-(4-methyl- piperazin-1-yl)-4-oxo-4H-chromen-2-yl]-methanoyl}-amino)-phenyl]- piperazine-1-caxboxylic acidcyclopentylamide 51

6-Methoxy-8-(4-methyl-piperazin-1- yl)-4-oxo-4H-chromene-2-carboxylicacid {4-[4-(1-pyrrolidin-1-yl- methanoyl)-piperazin-1-yl]- phenyl}-amide52

6-Methoxy-8-(4-methyl-piperazin-1- yl)-4-oxo-4H-chromene-2-carboxylicacid {4-(4-(propane-2-sulfonyl)- piperazin-1-yl]-phenyl}-amide 53

6-Methoxy-8-(4-methyl-piperazin-1- yl)-4-oxo-4H-chromene-2-carboxylicacid {4-[4-(2-methyl-propanoyl)- piperazin-1-yl]-phenyl}-amide 54

6-Methoxy-8-(4-methyl-piperazin-1- yl)-4-oxo-4H-chromene-2-carboxylicacid {4-(4-(1-morpholin-4-yl- methanoyl)-piperazin-1-yl]- phenyl}-amide55

6-Fluoro-8-(4-methyl-piperazin-1-yl)-4- oxo-4H-chromene-2-carboxylicacid (4- morpholin-4-yl-phenyl)-amide 56

6-Fluoro-8-(4-methyl-piperazin-1-yl)-4- oxo-4H-chromene-2-carboxylicacid [4-(4- methanesulfonyl-piperazin-1-yl)-phenyl]- amide 57

6-Fluoro-8-(4-methyl-piperazin-1-yl)-4- oxo-4H-chromene-2-carboxylicacid [4-(4- acetyl-piperazin-1-yl)-phenyl]-amide 58

6-Fluoro-8-(4-methyl-piperazin-1-yl)-4- oxo-4H-chromene-2-carboxylicacid (3- chloro-4-morpholin-4-yl-phenyl)-amide 59

6-Fluoro-8-(4-methyl-piperazin-1-yl)-4- oxo-4H-chromene-2-carboxylicacid (3- fluoro-4-morpholin-4-yl-phenyl)-amide 60

6-Fluoro-8-(4-methyl-piperazin-1-yl)-4- oxo-4H-chromene-2-carboxylicacid (3- cyano-4-morpholin-4-yl-phenyl)-amide 61

6-Fluoro-8-(4-methyl-piperazin-1-yl)-4- oxo-4H-chromene-2-carboxylicacid [4-(1- morpholin-4-yl-methanoyl)-phenyl]-amide 62

6-Methyl-8-(4-methyl-piperazin-1-yl)-4- oxo-4H-chromene-2-carboxylicacid (4- morpholin-4-yl-phenyl)-amide 63

6-Methyl-8-(4-methyl-piperazin-1-yl)-4- oxo-4H-chromene-2-carboxylicacid [4-(1- morpholin-4-yl-methanoyl)-phenyl]-amide 64

6-Methyl-8-(4-methyl-piperazin-1-yl)-4- oxo-4H-chromene-2-carboxylicacid (3- fluoro-4-morpholin-4-yl-phenyl)-amide 65

6-Chloro-8-(4-methyl-piperazin-1-yl)-4- oxo-4H-chromene-2-carboxylicacid (4- morpholin-4-yl-phenyl)-amide 66

5-Methyl-8-(4-methyl-piperazin-1-yl)-4- oxo-4H-chromene-2-carboxylicacid (4- morpholin-4-yl-phenyl)-amide 67

5-Methoxy-8-(4-methyl-piperazin-1- yl)-4-oxo-4H-chromene-2-carboxylicacid (4-morpholin-4-yl-phenyl)- amide 68

6-Methoxy-8-(4-methyl-piperazin-1- yl)-4-oxo-4H-chromene-2-carboxylicacid {4-[4-(3-hydroxy-propanoyl)- piperazin-1-yl]-phenyl}-amide 69

4-[4-({1-[6-Fluoro-8-(4-methyl- piperazin-1-yl)-4-oxo-4H-chromen-2-yl]-methanoyl}-amino)-phenyl]- piperazine-1-carboxylic acid tert-butyl ester 70

4-[4-({1-[6-Fluoro-8-(4-methyl- piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acid (4-piperazin-1-yl- phenyl)-amide 71

6-Fluoro-8-(4-methyl-piperazin-1- yl)-4-oxo-4H-chromene-2-carboxylicacid [4-(4-ethane sulfonyl-piperazin- 1-yl)-phenyl]-amide 72

6-Fluoro-8-(4-methyl-piperazin-1- yl)-4-oxo-4H-chromene-2-carboxylicacid [4-(4-propionyl-piperazin-1-yl)- phenyl]-amide 73

6-Fluoro-8-(4-methyl-piperazin-1- yl)-4-oxo-4H-chromene-2- carboxylicacid {4-[4-(3-hydroxy- propanoyl)-piperazin-1-yl]-phenyl}- amide 74

N-[8-(4-Methyl-piperazin-1-yl)-4- oxo-4H-chromen-2-yl]-4-morpholin-4-yl-benzamide 75

8-(4-Methyl-piperazin-1-yl)- chroman-2-carboxylic acid(4-morpholin-4-yl-phenyl)-amide 76

(+)-8-(4-Methyl-piperazin-1-yl)- chroman-2-carboxylic acid (4-morpholin-4-yl-phenyl)-amide 77

(−)-8-(4-Methyl-piperazin-1-yl)- chroman-2-carboxylic acid (4-morpholin-4-yl-phenyl)-amide 78

racemic-8-(4-methyl-piperazin-1-yl)- 4-oxo-chroman-2-carboxylic acid (4-morpholin-4-yl-phenyl)-amide 79

8-(4-Methyl-piperazin-1-yl)-4-oxo- chroman-2-carboxylic acid (4-morpholin-4-yl-phenyl)-amide (faster running isomer) 80

8-(4-Methyl-piperazin-1-yl)-4-oxo- chroman-2-carboxylic acid (4-morpholin-4-yl-phenyl)-amide (slower running isomer). 81

4-[4-({1-[6-Fluoro-8-(4-methyl- piperazin-1-yl)-4-oxo-4H-chromen-2-yl]-methanoyl}-amino)-phenyl]- piperazine-1-carboxylic acid ethylamide82

6-Methoxy-8-(4-methyl- [1,4]diazepan-1-yl)-4-oxo-4H-chromene-2-carboxylic acid (4- morpholin-4-yl-phenyl)-amide 83

6-Ethoxy-8-(4-methyl-piperazin-1- yl)-4-oxo-4H-chromene-2- carboxylicacid (4-morpholin-4-yl- phenyl)-amide 84

6-Ethoxy-8-(4-methyl-piperazin-1- yl)-4-oxo-4H-chromene-2- carboxylicacid [4-(4-propionyl- piperazin-1-yl)-phenyl]-amide 85

6-Methoxy-4-oxo-8-piperazin-1-yl-4H- chromene-2-carboxylic acid (4-morpholin-4-yl-phenyl)-amide 86

6-Hydroxy-8-(4-methyl-piperazin-1-yl)- 4-oxo-4H-chromene-2-carboxylicacid (4- morpholin-4-yl-phenyl)-amide 87

6-Methoxy-8-(4-methyl-[1,4]diazepan-1 yl)-4-oxo-1,4-dihydro-quinoline-2-carboxylic acid (4-morpholin-4-yl- phenyl)-amide 88

6-Methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-1,4-dihydro-quinoline-2-carboxylic acid(4-morpholin-4-yl-phenyl)-amide 89

6-Methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-1,4-dihydro-quinoline-2-carboxylic acid[4-(4-propionyl-piperazin-1-yl)- phenyl]-amide 90

6-Fluoro-8-(4-methyl-piperazin-1-yl)-4-oxo-1,4-dihydro-quinoline-2-carboxylic acid(4-morpholin-4-yl-phenyl)-amide 91

6-Fluoro-8-(4-methyl-piperazin-1-yl)-4-oxo-1,4-dihydro-quinoline-2-carboxylic acid[4-(4-propionyl-piperazin-1-yl)- phenyl]-amide 92

8-[(2-Dimethylamino-ethyl)-methyl- amino]-6-methoxy-4-oxo-1,4-dihydro-quinoline-2-carboxylic acid (4- morpholin-4-yl-phenyl)-amide 93

8-[(3-Dimethylamino-propyl)-methyl- amino]-6-methoxy-4-oxo-1,4-dihydro-quinoline-2-carboxylic acid (4- morpholin-4-yl-phenyl)-amide 94

8-((3R)-(+)-3-Dimethylamino-pyrrolidin-1-yl)-6-methoxy-4-oxo-1,4-dihydro- quinoline-2-carboxylic acid (4-morpholin-4-yl-phenyl)-amide 95

8-((3S)-(−)-3-Dimethylamino-pyrrolidin-1-yl)-6-methoxy-4-oxo-1,4-dihydro- quinoline-2-carboxylic acid (4-morpholin-4-yl-phenyl)-amide 96

6-Methoxy-8-[methyl-(1-methyl- pyrrolidin-3-yl)-amino]-4-oxo-1,4-dihydro-quinoline-2-carboxylic acid (4- morpholin-4-yl-phenyl)-amide 97

8-[Ethyl-(1-ethyl-pyrrolidin-3-yl)-amino]-6-methoxy-4-oxo-1,4-dihydro-quinoline-2- carboxylic acid(4-morpholin-4-yl- phenyl)-amide 98

4-dimethylamino-6-methoxy-8-(4-methyl-piperazin-1-yl)-quinoline-2-carboxylic acid(4-morpholin-4-yl-phenyl)-amide 99

6-methoxy-4-methylamino-8-(4-methyl-piperazin-1-yl)-quinoline-2-carboxylic acid(4-morpholin-4-yl-phenyl)-amide 100

6-fluoro-4-methoxy-8-(4-methyl- piperazin-1-yl)-quinoline-2-carboxylicacid (4-morpholin-4-yl-phenyl)-amide 101

6-Fluoro-4-oxo-8-piperazin-1-yl-4H- chromene-2-carboxylic acid(4-morpholin- 4-yl-phenyl)-amide

The following reference examples illustrate the making of intermediatesin the synthesis of the compounds of the present invention, and are notintend to limit the invention in any manner.

REFERENCE EXAMPLE 1 Preparation of Reference Example 18-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic AcidHydrochloride REFERENCE EXAMPLE 1a(E,Z)-2-(2-Bromo-phenoxy)-but-2-enedioic Acid Diethyl Ester

Diethyl acetylenedicarboxylate (20 ml, 0.162 mol) was added to2-bromophenol (28 g, 0.162 mol), in anhydrous 2-propanol (60 ml)followed by the addition of a catalytic amount of tetrabutylammoniumfluoride (0.5 ml, 1.0 M in THF). The solution was stirred at roomtemperature four hours and was then heated to reflux for one hour. Themixture was cooled to room temperature, then concentrated under vacuumto an oil (51 g=91%).

REFERENCE EXAMPLE 1b (E,Z)-2-(2-Bromo-phenoxy)-but-2-enedioic Acid

(E,Z)-2-(2-Bromo-phenoxy)-but-2-enedioic acid diethyl ester (51 g, 148mmol) as prepared in Reference Example 1a was suspended in ethanol (95ml) and a solution of sodium hydroxide (12.9 g, 0.323 mol) in water (95ml) was added. The solution was refluxed for 1 h to give a clear orangesolution. The mixture was cooled to room temperature and acidified with6 M HCl (50 ml). The mixture was then concentrated under vacuum and theresidue azeotroped (4×) with ethanol. The solid was filtered, washedwith water and dried to give(2Z)-2-(2-bromo-4-methoxyphenoxy)-2-butenedioic acid as a light orangesolid (24.3 g, 88% yield). This crude product was used without furtherpurification.

REFERENCE EXAMPLE 1c Ethyl-8-Bromo-4-oxo-4H-chromene-2-carboxylate

Sulfuric acid (95 mL) was added to crude(E,Z)-2-(2-Bromo-phenoxy)-but-2-enedioic acid as prepared in ReferenceExample 1b. After heating the mixture with a heat gun for 45 min anorange milky solution was obtained. This solution was slowly added torefluxing absolute ethanol (500 mL). After the addition, the reactionwas refluxed for 30 min then allowed to cool. Crystals started to formafter 20 min and the reaction was put in the refrigerator overnight. Thesolid was filtered, washed with cold ethanol/water 9:1 and dried to giveethyl 8-bromo-4-oxo-4H-chromene-2-carboxylate as an off-white solid(11.7 g, 24% yield, mp 124-126° C.).

REFERENCE EXAMPLE 1dEthyl-8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromenec-2-carboxylic Acid

Ethyl 8-bromo-4-oxo-4H-chromene-2-carboxylate as prepared in ReferenceExample 1c (Davies, Stephen et al., J. Chem. Soc. Perkin Trans I p 2597,1987) (3.0 g, 10.1 mmol) was azeotroped with anhydrous toluene then thewhite solid was dissolved in 100 mL anhydrous toluene and transferred tothe reaction vessel. The mixture was subjected to vacuum/argon (×2) andthe following were added in order (positive argon pressure):N-methylpiperazine (1.3 ml, 11.1 mmol), 2,2′-bis(diphenylphosphino)-1,1′-binaphthyl (0.75 g, 1.2 mmol,),tris(dibenzylideneacetone) dipalladium (0) (0.48 g, 0.5 mmol) thencesium carbonate (4.6 g, 14.1 mmol). The mixture was again subjected tovacuum/argon and was heated at 80° C. overnight.

The cooled reaction mixture was filtered through diatomaceous earth andthe toluene solution was applied directly to a 600 ml filter funnel(silica 230-400 mesh ASTM packed in ethyl acetate) and then washed withethyl acetate (2 l). The product was eluted with 5-8%methanol/chloroform and the desired was collected to give 2.5 g of aslightly impure orange yellow solid (mp 120-123° C.). The impure productwas chromatographed on a Waters Delta Prep 4000 using 1 PrepPakcartridge (Porasil 37-55 μm 125 Å) eluting with 3-5%methanol/chloroform. The product was collected and dried to give ethyl8-(4-methyl-1-piperazinyl)-4-oxo-4H-chromene-2-carboxylate as a yellowsolid (2.25 g, 70% yield mp 124-125° C.). GC/MS (EI, M+) m/z 316.

REFERENCE EXAMPLE 1e8-(4-methyl-1-piperazinyl)-4-oxo-4H-chromene-2-carboxylic AcidHydrochloride

Ethyl 8-(4-methyl-1-piperazinyl)-4-oxo-4H-chromene-2-carboxylate asprepared in Reference Example 1d (1.01 g. 3.19 mmol) was suspended in 6M HCl (60 ml) and to reflux for 1.5 h (after 20 min a clear solution wasobtained).

The reaction was allowed to cool. The solution was concentrated in vacuoand anhydrous toluene was added (×3) and the solution was againconcentrated in vacuo to give8-(4-methyl-1-piperazinyl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride as a yellow powder (1.02 g, quantitative yield). LC/MS(M+1) m/z 289.

REFERENCE EXAMPLE 2

Preparation of6-Methoxy-8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicAcid Hydrochloride REFERENCE EXAMPLE 2a Diethyl(2Z)-2-(2-bromo-4-methoxyphenoxy)-2-butenedioate

Ethyl acetylenedicarboxylate (17.8 ml, 0.145 mol) was added to2-bromo-4-methoxyphenol (Synlett p1241, 1997) (27.3 g, 0.134 mol), inanhydrous 2-propanol (55 ml) followed by the addition of a catalyticamount of tetrabutylammonium fluoride (0.4 ml, 1.0 M in THF). Thesolution was stirred at room temperature overnight and was then heatedto reflux for 30 min. Upon cooling a precipitate formed. The solutionwas cooled and filtered to give diethyl(2Z)-2-(2-bromo-4-methoxyphenoxy)-2-butenedioate as a yellow solid (29.9g, 62% yield). Note: the solid contains 10% of diethyl(2E)-2-(2-bromo-4-methoxyphenoxy)-2-butenedioate. GC/MS (EI, M+) m/z 344and 346.

REFERENCE EXAMPLE 2b (2Z)-2-(2-bromo-4-methoxyphenoxy)-2-butenedioicAcid

Diethyl (2Z)-2-(2-bromo-4-methoxyphenoxy)-2-butenedioate (29.9 g, 86.6mmol) as prepared in Reference Example 2a was suspended in ethanol (55ml) and a solution of sodium hydroxide (7.0 g, 0.175 mol) in water (55ml) was added. The solution was refluxed for 1 h to give a clear orangesolution. Most of the ethanol was removed in vacuo then 6 M HCl (50 ml)was added. The solid was filtered, washed with water and dried to give(2Z)-2-(2-bromo-4-methoxyphenoxy)-2-butenedioic acid as a light orangesolid (24.3 g, 88% yield).

REFERENCE EXAMPLE 2cEthyl-6-methoxy-8-bromo-4-oxo-4H-chromene-2-carboxylate

Sulfuric acid (50 ml) was added to(2Z)-2-(2-bromo-4-methoxyphenoxy)-2-butenedioic acid (24.3 g, 86.6 mmol;as prepared in Reference Example 2b above). After heating the mixturewith a heat gun for 5-10 min a clear deep brown solution was obtained.This solution was slowly added to refluxing absolute ethanol (250 ml).After the addition the reaction was refluxed for 30 min then allowed tocool. Crystals started to form after 20 min and the reaction was put inthe refrigerator overnight. The solid was filtered, washed with coldethanol/water 9:1 and dried to give ethyl8-bromo-6-methoxy-4-oxo-4H-chromene-2-carboxylate as an off-white solid(12.3 g, 50% yield, mp 159-161° C.).

REFERENCE EXAMPLE 2dEthyl-6-methoxy-8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylate

Ethyl 8-bromo-4-oxo-4H-chromene-2-carboxylate (9.2 g, 28.1 mmol), asprepared in Example 2c above, was azeotroped with anhydrous toluene thenthe white solid was dissolved in 300 ml anhydrous toluene in a 500 mLsingle-neck round bottom flask. The mixture was degassed by alternatingargon sparge and vacuum (3×), and the following were added in order:N-methylpiperazine (4.0 ml, 35.1 mmol), 2,2′-bis(diphenylphosphino)-1,1′-binaphthyl (1.05 g, 1.69 mmol,),tris(dibenzylideneacetone) dipalladium (0) (0.50 g, 0.56 mmol) thencesium carbonate (12.8 g, 39.3 mmol). The mixture was again degassed viaalternating argon sparge and vacuum and was heated at 80° C. for 17 h.Additional tris(dibenzylideneacetone) dipalladium (0) (0.10 g, 0.11mmol) and 2,2′-bis (diphenylphosphino)-1,1′-binaphthyl (0.20 g, 0.32mmol,) was added and the reaction was stirred at 80° C. for another 55 hat which time the conversion was essentially complete.

The cooled reaction mixture was diluted with tetrahydrofuran (250 mL),filtered and concentrated under vacuum. The residue was purified bychromatography on a silica column eluted with 2-5% methanol/chloroformand the desired fractions were collected and concentrated under vacuumand the residue triturated with methylene chloride to give 7.4 g (76%)of a yellow powder.

REFERENCE EXAMPLE 2e6-Methoxy-8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicAcid

Ethyl-6-methoxy-8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylate(1.0 g. 2.89 mmol), as prepared in Reference Example 2d above, wassuspended in 6 M HCl (60 ml) and methanol (10 mL) and warmed to refluxfor 3.0 h. The reaction was allowed to cool. The solution wasconcentrated in vacuo and anhydrous toluene was added (×3) and thesolution was again concentrated in vacuo. The residue was dried undervacuum (17 h) to yield6-methoxy-8-(4-methyl-1-piperazinyl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride as a yellow powder (1.0 g, quantitative yield).

REFERENCE EXAMPLE 3

6-Fluoro-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic AcidHydrochloride REFERENCE EXAMPLE 3a Diethyl(EZ)-2-(2-bromo-4-fluorophenoxy)-2-butenedioate

This compound was synthesized from 2-bromo-4-fluorophenol anddiethylacetylenedicarboxylate, using the same synthetic procedures andthe same stoichiometry as demonstrated in Reference Example 1a above.

REFERENCE EXAMPLE 3b (EZ)-2-(2-Bromo-4-fluorophenoxy)-2-butenedioic Acid

This compound was synthesized from diethyl(EZ)-2-(2-bromo-4-fluorophenoxy)-2-butenedioate, as prepared inReference Example 3a above, using the same synthetic procedures and thesame stoichiometry as demonstrated in Reference Example 1b above.

REFERENCE EXAMPLE 3cEthyl-6-fluoro-8-bromo-4-oxo-4H-chromene-2-carboxylate

This compound was synthesized from(EZ)-2-(2-bromo-4-fluorophenoxy)-2-butenedioic acid, as prepared inReference Example 3b above, using the same synthetic procedures and thesame stoichiometry as demonstrated in Reference Example 1c above.

REFERENCE EXAMPLE 3dEthyl-6-fluoro-8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylate

This compound was synthesized fromethyl-6-fluoro-8-bromo-4-oxo-4H-chromene-2-carboxylate, as prepared inReference Example 3c above, using the same synthetic procedures and thesame stoichiometry as demonstrated in Reference Example 1d above.

REFERENCE EXAMPLE 3e6-Fluoro-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic AcidHydrochloride

This compound was synthesized starting fromethyl-6-methoxy-8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylate,as prepared in Example 3d, using the same synthetic procedures and thesame stoichiometry as demonstrated in Reference Example 1e above.

REFERENCE EXAMPLE 4

Preparation6-Methyl-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic AcidHydrochloride REFERENCE EXAMPLE 4a Diethyl(E,Z)-2-(2-bromo-4-methylphenoxy)-2-butenedioate

2-Bromo-4-methyl phenol (10 mL, 83 mmol) was dissolved in diethyl ether(90 mL). To this was added dropwise triethyl amine (13.7 mL, 98 mmol)followed by dimethyl acetylene dicarboxylate (11.2 mL, 91 mmol). Theresulting mixture was stirred overnight at room temperature. Thereaction was worked up by adding diethyl ether (200 mL) andtetrahydrofuran (50 mL) and washing the resulting mixture with 1N HCl(200 mL), water (200 mL) and brine (100 mL). The organic phase was thendried (Na₂SO₄), filtered and concentrated to a red-brown oil which wasused without further purification.

REFERENCE 4b (2E,Z)-2-(2-Bromo-4-fluorophenoxy)-2-butenedioic Acid

This compound was synthesized from diethyl(E,Z)-2-(2-bromo-4-methylphenoxy)-2-butenedioate, as prepared inReference Example 4a above, using the same synthetic procedures and thesame stoichiometry as demonstrated in Example 1b above.

REFERENCE EXAMPLE 4cEthyl-6-methyl-8-bromo-4-oxo-4H-chromene-2-carboxylate

This compound was synthesized from(2Z)-2-(2-bromo-4-methylphenoxy)-2-butenedioic acid, as prepared inReference Example 4b above, and using the same synthetic procedures andthe same stoichiometry as demonstrated in Reference Example 1c above.

REFERENCE EXAMPLE 4dEthyl-6-methyl-8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylate

This compound was synthesized fromethyl-6-methyl-8-bromo-4-oxo-4H-chromene-2-carboxylate, as prepared inReference Example 4c above, using the same synthetic procedures and thesame stoichiometry as demonstrated in Reference Example 1d above.

REFERENCE EXAMPLE 4e6-Methyl-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic AcidHydrochloride

This compound was synthesized starting withethyl-6-methyl-8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylate,as prepared in Reference Example 4d, using the same synthetic proceduresand the same stoichiometry as demonstrated in Reference Example 1eabove.

REFERENCE EXAMPLE 5

Preparation of6-Chloro-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic AcidHydrochloride REFERENCE EXAMPLE 5a Diethyl(E,Z)-2-(2-bromo-4-chlorophenoxy)-2-butenedioate

This compound was prepared from 2-bromo-4-chloro phenol and dimethylacetylenedicarboxylate by the same synthetic procedures and in the samestoichiometry as the preparation described in Reference Example 4a.

REFERENCE EXAMPLE 5b (2E,Z)-2-(2-Bromo-4-chlorophenoxy)-2-butenedioicAcid

This compound was synthesized from diethyl(E,Z)-2-(2-bromo-4-chlorophenoxy)-2-butenedioate, as prepared inReference Example 5a above, as using the same synthetic procedures andthe same stoichiometry as demonstrated in Reference Example 1b above.

REFERENCE EXAMPLE 5cEthyl-6-chloro-8-bromo-4-oxo-4H-chromene-2-carboxylate

This compound was synthesized from(2E,Z)-2-(2-bromo-4-chlorophenoxy)-2-butenedioic acid, as prepared inReference Example 5b above, using the same synthetic procedures and thesame stoichiometry as demonstrated in Example 1c above.

REFERENCE EXAMPLE 5dEthyl-6-chloro-8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylate

This compound was synthesized fromethyl-6-chloro-8-bromo-4-oxo-4H-chromene-2-carboxylate, as prepared inReference Example 5c above, using the same synthetic procedures and thesame stoichiometry as demonstrated in Example 1d above.

REFERENCE EXAMPLE 5e6-Chloro-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic AcidHydrochloride

This compound was synthesized starting withethyl-6-chloro-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylate,prepared in Reference Example 5d above, using the same syntheticprocedures and the same stoichiometry as demonstrated in ReferenceExample 1e above.

REFERENCE EXAMPLE 6

Preparation of5-Methyl-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic AcidHydrochloride REFERENCE EXAMPLE 6a Diethyl(E,Z)-2-(2-chloro-5-methylphenoxy)-2-butenedioate

This compound was prepared from 2-chloro-5-methylphenol and dimethylacetylenedicarboxylate by the same synthetic procedures and in the samestoichiometry as the preparation described in Reference Example 1a.

REFERENCE EXAMPLE 6b (2E,Z)-2-(2-chloro-5-methylphenoxy)-2-butenedioicAcid

This compound was synthesized from diethyl(E,Z)-2-(2-chloro-5-methylphenoxy)-2-butenedioate, as prepared inReference Example 6a above, using the same synthetic procedures and thesame stoichiometry as demonstrated in Reference Example 1b above.

REFERENCE EXAMPLE 6cEthyl-5-methyl-8-chloro-4-oxo-4H-chromene-2-carboxylate

This compound was synthesized from(2Z)-2-(2-chloro-5-methylphenoxy)-2-butenedioic acid, as prepared inReference example 6b, using the same synthetic procedures and the samestoichiometry as demonstrated in Reference Example 1c above.

REFERENCE EXAMPLE 6dEthyl-5-methyl-8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylate

Ethyl 5-methyl-8-chloro-4-oxo-4H-chromene-2-carboxylate (1.0 g, 3.6mmol) as prepared in Reference Example 6c above, was azeotroped withanhydrous toluene then the white solid was dissolved in 100 ml anhydroustoluene in a 250 mL single-neck round bottom flask. The mixture wasdegassed by alternating argon sparge and vacuum (3×), and the followingwere added in order: N-methylpiperazine (0.6 ml, 5.37 mmol),(2′-dicyclohexylphosphanyl-biphenyl-2-yl)-dimethyl-amine (JACS 1998,120, p9722) (40 mg, 0.1 mmol,), tris(dibenzylideneacetone) dipalladium(0) (66 mg, 0.072 mmol) then cesium carbonate (1.6 g, 5.37 mmol). Themixture was again degassed via alternating argon sparge and vacuum andwas heated at 80° C. for 17 h. Additional tris(dibenzylideneacetone)dipalladium (0) (66 mg, 0.072 mmol) and(2′-dicyclopentylphosphanyl-biphenyl-2-yl)-dimethyl-amine (40 g, 0.1mmol,) were added and the reaction was stirred at 80° C. for anotherfour days at which time the conversion was still only about 50% completeby HPLC. Tetrahydrofuran (100 mL) was added, and the combined mixturewas filtered, concentrated under vacuum and purified by chromatographyon silica eluted with 2.5% methanol in chloroform. The desired fractionswere concentrated under vacuum to yield a yellow powder (250 mg=21%).

REFERENCE EXAMPLE 6e5-Methyl-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic AcidHydrochloride

This compound was synthesized starting withethyl-5-methyl-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylate,as prepared in Reference Example 6d, and using the same syntheticprocedures and the same stoichiometry as demonstrated in Example 1eabove.

REFERENCE EXAMPLE 7

Preparation of5-Methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicAcid Hydrochloride REFERENCE EXAMPLE 7a(E,Z)-2-(2-Bromo-5-methoxyphenoxy)-2-butenedioate

This compound was prepared from 2-bromo-5-methoxyphenol and dimethylacetylenedicarboxylate by the same synthetic procedures and in the samestoichiometry as the preparation described in Reference Example 1a.

REFERENCE EXAMPLE 7b (E,Z)-2-(2-Bromo-5-methoxyphenoxy)-2-butenedioicAcid

This compound was synthesized from diethyl(E,Z)-2-(2-bromo-5-methoxyphenoxy)-2-butenedioate, as prepared inReference Example 7a, using the same synthetic procedures and the samestoichiometry as demonstrated in Reference Example 1b above.

REFERENCE EXAMPLE 7cEthyl-5-methoxy-8-bromo-4-oxo-4H-chromene-2-carboxylate

This compound was synthesized from(E,Z)-2-(2-bromo-5-methoxyphenoxy)-2-butenedioic acid, as prepared inReference Example 7b above, using the same synthetic procedures and thesame stoichiometry as demonstrated in Reference Example 1c above.

REFERENCE EXAMPLE 7dEthyl-5-methoxy-8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylate

This compound was synthesized fromethyl-5-methoxy-8-bromo-4-oxo-4H-chromene-2-carboxylate, as prepared inReference Example 7c above, using the same synthetic procedures and thesame stoichiometry as demonstrated in Reference Example 1d above.

REFERENCE EXAMPLE 7e5-Methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicAcid Hydrochloride

This compound was prepared fromethyl-5-methoxy-8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylate,as prepared in Reference Example 7d above, using the same method as thepreparation in 1e.

REFERENCE EXAMPLE 8

Preparation of 1-(6-Piperazin-1-yl-2,3-dihydro-indol-1-yl)-ethanoneREFERENCE EXAMPLE 8a1-[5-(4-Benzyl-piperazin-1-yl)-2,3-dihydro-indol-1-yl]-ethanone

1-acetyl-5-bromoindoline (3.0 g, 12.5 mmol) was dissolved in toluene (60mL). To this was added, sodium t-butoxide (1.68 g, 17.5 mmol),N-benzylpiperazine (2.4 mL, 13.8 mmol), S-BINAP (0.93 g, 1.5 mmol) andPd₂(dba)₃ (0.46 g, 0.5 mmol). The mixture was degassed via three cyclesof vacuum and nitrogen sparge and then stirred at 95° C. until GCanalysis confirmed that the reaction was complete (1 h). The mixture wasdiluted with ethyl acetate (150 mL), washed with water and extractedwith 2N HCl (2×100 mL). The combined aqueous extract was basified withconcentrated ammonium hydroxide and extracted with ethyl acetate (2×100mL). The combined organic extract was dried (MgSO₄) and concentrated toyield a solid (2.7 g) which was purified by chromatography to yield awhite solid (1.81 g, 43%). Mp=150.5-152.8° C.

REFERENCE EXAMPLE 8b1-(6-piperazin-1-yl-2,3-dihydro-indol-1-yl)-ethanone

1-[5-(4-Benzyl-piperazin-1-yl)-2,3-dihydro-indol-1-yl]-ethanone (0.37 g,1.1 mmol), as prepared in Reference Example 8a above, was dissolved inmethanol (5 mL). Pd/C (90 mg, 10%) and ammonium formate (0.9 g, 14 mmol)was added and the resulting mixture was heated to 65° C. for two hours.The mixture was filtered and the filter cake washed with hot methanol.The combined filtrate was concentrated to yield the desired product(0.26 g, 90%).

REFERENCE EXAMPLE 9

Preparation of 2-chloro-5-piperazin-1-yl benzonitrile REFERENCE EXAMPLE9a 3-Cyano-4-chloroaniline

2-Chloro-5-nitrobenzonitrile (25 g, 137 mmol) was dissolved in ethanol(275 mL). Stannous chloride dihydrate (154.5 g, 0.685 M) was added andthe mixture stirred at 70° C. for 30 min. The mixture was then cooled toroom temperature and poured into crushed ice. The mixture was made basicwith solid sodium hydroxide. This mixture was extracted with ethylacetate (3×100 mL). The extracts were combined, washed with brine, dried(MgSO₄), concentrated and the residue dried under vacuum andrecrystallized from ethanol to yield light brown needles (10.6 g, 51%).

REFERENCE EXAMPLE 9b 2-chloro-5-piperazin-1-yl Benzonitrile

3-Cyano-4-chloroaniline (10.1 g, 66 mmol), as prepared in ReferenceExample 9a, was dissolved in n-butanol (300 mL) bis-(2-chloroethyl)aminehydrochloride (23.2 g, 130 mmol) and potassium iodide (50 mg, catalytic)were added. The mixture was heated at reflux for three days, then cooledin a refrigerator overnight. A solid precipitate was collected byfiltration, washed with cold n-butanol and dried. The crude product wasdistributed between methylene chloride and 2N ammonium hydroxide. Theorganic layer was separated, dried (Na₂SO₄) and concentrated to yield alight yellow solid (9.1 g, 59%) which gave a single peak by GC and TLCanalysis.

REFERENCE EXAMPLE 10

Preparation of 4-[1,2,3]thiadiazol-5-yl-phenylamine

SnCl₂.H₂O (3.21 g, 5 eq) was added to a slurry of(5-(4-Nitrophenyl)-1,2,3-thiadiazole (Lancaster Synthesis) (0.59 g, 2.8mmol) in absolute EtOH (50 mL) and the reaction heated to 70° C. for 2h. The reaction was allowed to cool to room temperature and pour intosaturated NaHCO₃ and ice. The product was extracted with EtOAc (2×) thesolution dried (MgSO₄) and evaporated to dryness in vacuo to yield 0.47g of a light yellow solid mp 126-128° C.

REFERENCE EXAMPLE 11

Preparation of 1-[4-(4-Amino-phenyl)-piperazin-1-yl]-ethanone REFERENCEEXAMPLE 11a 4(4-Nitrophenyl)-1-acetylpiperazine

1-(4-Nitrophenyl)piperazine (2.5 g, 12.1 mmol) was dissolved indichloromethane (100 ml). Triethylamine (2.0 ml, 14.5 mmol) was addedand the reaction was cooled to 0° C. Acetic anhydride (1.25 ml, 13.3mmol) was added dropwise and the reaction was stirred at 0° C. for 1 h.Saturated sodium bicarbonate was added and the reaction was extracted(×3) with dichloromethane, dried (MgSO₄), filtered and concentrated invacuo to give 4-(4-nitrophenyl)-1-acetylpiperazine as a yellow solid(3.01 g,). GC/MS (EI, M+) m/z=249.

REFERENCE EXAMPLE 11b 1-[4-(4-Amino-phenyl)-piperazin-1-yl]-ethanone

4-(4-Nitrophenyl)-1-acetylpiperazine (3.0 g, 12.0 mmol), as prepared inReference Example 11a above, was mixed in methanol (100 ml) and 2 Mammonia in methanol (50 ml) and 10% palladium on carbon (300 mg) wasadded. The mixture was hydrogenated on a Paar apparatus (50 psi) for 1.5h.

The reaction was allowed to cool, the catalyst was filtered and thesolution was concentrated in vacuo. The crude solid was recrystallizedfrom ethyl acetate to give 4-(4-acetyl-1-piperazinyl)benzenamine as alight purple solid (1.86 g, 70% yield, mp 149.5-150.5° C.). GC/MS (EI,M+) m/z=219.

REFERENCE EXAMPLE 12

Preparation of 4-(4-methanesulfonyl-piperazin-1-yl)-phenylamineREFERENCE 12a 4-(4-Nitrophenyl)-1-methylsulfonylpiperazine

1-(4-Nitrophenyl)piperazine (2.79 g, 13.5 mmol) was dissolved indichloromethane (100 ml). Triethylamine (2.25 ml, 16.2 mmol) was addedand the reaction was cooled to 0° C. Methanesulfonyl chloride (1.15 ml,14.9 mmol) was added dropwise and the reaction was stirred at 0° C. for1 h. Saturated sodium bicarbonate was added and the reaction wasextracted (×3) with dichloromethane, dried (MgSO₄), filtered andconcentrated in vacuo to give4-(4-nitrophenyl)-1-methylsulfonylpiperazine as a yellow solid (3.83 g,quantitative yield). GC/MS (EI, M+) m/z=285.

REFERENCE EXAMPLE 12b 4-(4-methanesulfonyl-piperazin-1-yl)-phenylamine

4-(4-Nitrophenyl)-1-methylsulfonylpiperazine (3.83 g, 13.4 mmol), asprepared in Reference Example 12a above, was mixed in methanol (100 ml)and 2 M ammonia in methanol (50 ml) and 10% palladium on carbon (400 mg)was added. The mixture was hydrogenated on a Paar apparatus (50 psi) for3 h.

The reaction was allowed to cool, the catalyst was filtered, washed withmethanol then washed with chloroform. The chloroform portion contained aminor amount of the desired but looked purer. The chloroform portion wasconcentrated in vacuo and was recrystallized ethyl acetate to give4-[4-(methylsulfonyl)-1-piperazinyl]benzenamine as a shiny brown solid(0.94 g, 27% yield, mp 192-193° C.). GC/MS (EI, M+) m/z=255.

REFERENCE EXAMPLE 13

Preparation of 4-Thiomorpholin-4-yl-phenylamine REFERENCE EXAMPLE 13a4-(4-Nitro-phenyl)-thiomorpholine

4-Fluoronitrobenzene (3.0 g, 21.3 mmol) was dissolved in toluene (25mL). Thiomorpholine (2.4 mL, 23.4 mmol) was added and the mixturestirred overnight at 100° C. At 17 h, the mixture was distributedbetween ethyl acetate (100 mL) and saturated sodium bicarbonate (50 mL).The organic layer was separated, dried (Na₂SO₄), filtered andconcentrated under vacuum. The residue was triturated with hexane toyield a bright yellow solid.

REFERENCE EXAMPLE 13b 4-Thiomorpholin-4-yl-phenylamine

4-(4-Nitro-phenyl)-thiomorpholine(3.0g, 13.4 mmol), as prepared inReference Example 13a above, was dissolved in ethanol (250 mL) and 10%palladium on carbon (250 mg) was added. This mixture was shaken on aParr hydrogenator for 3 h. The reaction mixture was then filteredthrough diatomaceous earth and concentrated under vacuum. The residuewas triturated with hexane to yield an gray solid (2.1 g).

REFERENCE EXAMPLE 14

Preparation of 1-(4-Amino-phenyl)-1-morpholin-4-yl-methanone REFERENCEEXAMPLE 14a 1-Morpholin-4-yl-1-(4-nitro-phenyl)-methanone

4-Nitrobenzoyl chloride (5 g, 27 mmol) in tetrahydrofuran (10 mL) wasadded slowly to a solution of morpholine (5g, 88 mmol) and triethylamine(2.7 g, 27 mmol) in tetrahydrofuran (50 mL), and stirred at roomtemperature for four hours. Ethyl acetate (200 mL) was added to themixture and the combined mixture was washed with water (25 mL), 1N HCl(25 mL), water (25 mL), saturated sodium bicarbonate (25 mL), water (25mL) and brine (25 mL). The mixture was dried (Na2SO4), filtered andconcentrated under vacuum and the residue used without furtherpurification.

REFERENCE EXAMPLE 14b 1-(4-Amino-phenyl)-1-morpholin-4-yl-methanone

This compound was prepared from1-morpholin-4-yl-1-(4-nitro-phenyl)-methanone as prepared in ReferenceExample 13b.

REFERENCE EXAMPLE 15

Preparation of 5-Amino-2-morpholin-4-yl-benzonitrile REFERENCE EXAMPLE15a 2-Morpholin-4-yl-5-nitro-benzonitrile

3-Cyano-4-fluoronitrobenzene (3.3 g, 19.9 mmol) was dissolved in ethylacetate (10 mL). Morpholine (2.2 mL, 25 mmol), andN,N-diisopropylethylamine (3.5 mL, 20 mmol) were added and the mixturestirred overnight at room temperature. At 17 h, additional ethyl acetate(150 mL) was added and the combined mixture was washed with water (50mL) and brine (50 mL), dried (Na₂SO₄), filtered and concentrated undervacuum. The residue was used without further purification.

REFERENCE EXAMPLE 15b 5-Amino-2-morpholin-4-yl-benzonitrile

This compound was prepared from 2-Morpholin-4-yl-5-nitro-benzonitrile(as prepared in Reference Example 15a above), as prepared in ReferenceExample 13b.

REFERENCE EXAMPLE 16

Preparation of 3-fluoro-4-morpholin-4-yl-phenylamine REFERENCE EXAMPLE16a 4-(2-Fluoro-4-nitro-phenyl)-morpholine

3,4-Difluoronitrobenzene (3.7 g, 23.2 mmol) was dissolved in ethylacetate (10 mL). Morpholine (2.2 mL, 25 mmol), andN,N-diisopropylethylamine (4 mL, 23 mmol) were added and the mixturestirred overnight at room temperature. At 17 h, additional ethyl acetate(150 mL) was added and the combined mixture was washed with water (50mL) and brine (50 mL), dried (Na₂SO₄), filtered and concentrated undervacuum. The residue was used without further purification.

REFERENCE EXAMPLE 16b 3-Fluoro-4-morpholin-4-yl-phenylamine

This compound was prepared from 4-(2-fluoro-4-nitro-phenyl)-morpholine,(as prepared in Reference Example 16a above) as prepared in ReferenceExample 13b.

REFERENCE EXAMPLE 17

Preparation of 4-(4-Amino-phenyl)-piperazine-1-carboxylic AcidTert-Butyl Ester REFERENCE EXAMPLE 17a4-(4-Nitro-phenyl)-piperazine-1-carboxylic Acid Tert-Butyl Ester

4-Fluoronitrobenzene (4.8 g, 34 mmol) was dissolved in ethyl acetate (25mL). Piperazine-1-carboxylic acid tert-butyl ester (6.7 g, 36 mmol) andN,N-diisopropylethylamine (6.3 mL, 36 mmol) were added and the mixturewas stirred at 65° C. for five days and cooled to room temperature.Ether (100 mL) was added and the combined mixture was washed with water(25 mL) and brine (25 mL), dried (Na2SO4), filtered and concentratedunder vacuum. The residue was triturated with hexane to yield a brightyellow solid (8 g, 77%).

REFERENCE EXAMPLE 17b 4-(4-Amino-phenyl)-piperazine-1-carboxylic AcidTert-Butyl Ester

4-(4-Amino-phenyl)-piperazine-1-carboxylic acid tert-butyl ester wasprepared from 4-(4-Nitro-phenyl)-piperazine-1-carboxylic acid tert-butylester, (as prepared in Reference Example 17a) as prepared in ReferenceExample 13b.

REFERENCE EXAMPLE 18

Preparation of 3-Morpholin-4-yl-phenylamine REFERENCE EXAMPLE 18a4-(3-Nitro-phenyl)-morpholine

3-Fluoronitrobenzene (10 g, 71 mmol) was dissolved in acetonitrile (100mL). Morpholine (30 mL, 350 mmol) was added and the mixture was reacted18 h at 150° C./80 psi in a pressure reactor. The reaction was cooled toroom temperature, concentrated under vacuum and 5g of the total mixturewas purified by column chromatography on silica eluted with CH₂Cl₂. Theproduct (3.6 g) was isolated as a bright yellow oil.

REFERENCE EXAMPLE 18b 3-Morpholin-4-yl-phenylamine

3-Morpholin-4-yl-phenylamine was prepared from4-(3-Nitro-phenyl)-morpholine, (as prepared in Reference Example 18a),as prepared in Reference Example 13b.

REFERENCE EXAMPLE 19

Preparation of 2-[4-(4-amino-phenyl)-piperazin-1-yl]-ethanol REFERENCEEXAMPLE 19a 2[4-(4-nitrophenyl)piperazine-1-yl]-ethanol

2[4-(4-nitrophenyl)piperazine-1-yl]-ethanol is prepared fromcommercially available 4-fluoronitrobenzene (Aldrich) and commerciallyavailable N-(2-hydroxyethyl)piperazine (Aldrich) via the same procedureas described in Reference Example 13a above.

REFERENCE EXAMPLE 19b 2-[4-(4-amino-phenyl)-piperazin-1-yl]-ethanol

2-[4-(4-amino-phenyl)-piperazin-1-yl]-ethanol is prepared by catalytichydrogenation of 2[4-(4-nitrophenyl)piperazine-1-yl]-ethanol (preparedas in Reference Example 19a) as described in Reference Example 13b.

REFERENCE EXAMPLE 20

Preparation of 4-Morpholin-4-yl-phenylamine

4-(4-Nitrophenyl)morpholine (10.3 g, 49.5 mmol;) (Lancaster Synthesis)was suspended in methanol (130 ml) and 2 M ammonia in methanol (70 mL)and 5% palladium on carbon (100 mg) was added. The mixture washydrogenated on a Paar apparatus (50 psi) for 1 h. The reaction wasallowed to cool, the catalyst was filtered and the solution wasconcentrated in vacuo. The crude solid was recrystallized from ethylacetate/hexane to give 4-(4-morpholinyl)aniline as a light purple solid(6.2 g, 70% yield, mp 132-133° C.). GC/MS (EI, M+) m/z=178.

REFERENCE EXAMPLE 21

Preparation of 4-Amino-3-hydroxyphenylmorpholine

4-Nitro-3-hydroxyphenylmorpholine (Maybridge Chemical) (3.34 g, 14.9mmol) was dissolved in 59 ml of ethanol at 30° C. The mixture wasstirred at 25° C. and treated with tin (II) chloride dihydrate (16.8grams, 74.5 mmol) with stirring. The yellow suspension was heated toreflux over a 30 minute period. TLC showed reaction progress overseveral hours. The mixture was refluxed for 18 hours, cooled to roomtemperature, and concentrated to remove most of the ethanol to give ayellow slurry. The mixture was treated with saturated aqueous sodiumbicarbonate until it was basic. The mixture was extracted with ethylacetate, filtered, and the organic layer was separated. The aqueouslayer was extracted twice more with ethyl acetate. The extracts werecombined, dried over magnesium sulfate, filtered, and concentrated togive 1.02 grams of a purple solid. Proton NMR and CI mass spectralanalyses were consistent for the desired product (m/z=195 base peak bypositive ion CI and m/z=193 base peak by negative ion CI).

REFERENCE EXAMPLE 22

Preparation of6-Methoxy-8-(4-methyl-[1,4]diazepan-1-yl)-4-oxo-4H-chromene-2-carboxylicAcid REFERENCE EXAMPLE 22a6-Methoxy-8-(4-methyl-[1,4]diazepan-1-yl)-4-oxo-4H-chromene-2-carboxylicAcid Ethyl Ester

Into a 250 mL 3 neck round bottom flask equipped with a refluxcondenser, nitrogen inlet and magnetic stirrer is placed 1.5 g (4.59mmol, 1.0 equiv.) of 8-Bromo-6-methoxy-4-oxo-4H-chromene-2-carboxylicacid ethyl ester (Reference Example 2c), 84 mg (0.092 mmol, 0.02 equiv.)of tris dibenzylidineacetone dipalladium, 342 mg (0.55 mmol, 0.12equiv.) of racemic 2,2′-bis(diphenylphosphino)-1,1′-binapthyl and 2 g of4 A molecular sieves. To this suspension is added 150 mL of dry toluene.To the stirred suspension is then added 628 mg, 684 μL, (5.50 mmol, 1.2equiv.) of 1-methylhomopiperazine, followed by 2.05 g (6.3 mmol, 1.4equiv.) of cesium carbonate. The mixture is then heated to 80° C. for 3days. At the end of this time completion was monitored by LC/MS analysisof an aliquot. When the reaction was determined to be complete it wascooled to room temperature then filtered through a plug of diatomaceousearth with toluene washing to remove solid by products. Purification byflash chromatography, using a gradient of 5 to 20% methanol in methylenechloride as eluent, yielded 1.0 g, (60%) of the desired product.

Mass Spec.: calc. for [C₁₉H₂₄N₂O₅+H]⁺ Theor. m/z=361; Obs. =361.

REFERENCE EXAMPLE 22b6-Methoxy-8-(4-methyl-[1,4]diazepan-1-yl)-4-oxo-4H-chromene-2-carboxylicAcid

Into a 125 mL erlenmeyer equipped with a magnetic stirrer is placed 319mg (0.89 mmol, 1.0 equiv.) of6-Methoxy-8-(4-methyl-[1,4]diazepan-1-yl)-4-oxo-4H-chromene-2-carboxylicacid ethyl ester. This material is dissolved in 30 mL of THF, then 30 mLof methanol are added. To this stirring solution is added 30 mL of awater containing 41 mg (0.97 mmol, 1.1 equiv.) of lithium hydroxide.This mixture is stirred at room temperature for 2 hr. Completion of thereaction is monitored by LC/MS, then 10 mL of 2N HCl is added. Thismixture is then concentrated, dried and triturated with ether to givethe product as the hydrochloride salt in quantitative yield.

Mass Spec.: calc. for [C₁₇H₂₀N₂O₅+H]⁺ Theor. m/z=333; Obs.=333.

REFERENCE EXAMPLE 23

Preparation of6-Ethoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carbonylChloride REFERENCE EXAMPLE 23a8-Bromo-6-hydroxy-4-oxo-4H-chromene-2-carboxylic Acid Ethyl Ester

The hydroxy compound, 8-Bromo-6-hydroxy-4-oxo-4H-chromene-2-carboxylicacid ethyl ester, is formed as a side product during the synthesis of8-Bromo-6-methoxy-4-oxo-4H-chromene-2-carboxylic acid ethyl ester. Itcan be separated from the crude methoxy compound by flash chromatographyusing a step gradient of 20% ethyl acetate in methylene chloride to thesame solvent containing 2% methanol. The hydroxy compound, which eluteslast, is concentrated to give the pure compound. Mass Spec.: calc. for[C₁₂H₉BrO₅+H]⁺ Theor. m/z=313, 315; Obs.=313, 315.

REFERENCE EXAMPLE 23b 8-Bromo-6-ethoxy-4-oxo-4H-chromene-2-carboxylicAcid Ethyl Ester

Into a 100 mL 3 neck round bottom flask equipped with a refluxcondenser, nitrogen inlet and magnetic stirrer is added 700 mg (2.24 mg,1.0 equiv.) of 8-Bromo-6-hydroxy-4-oxo-4H-chromene-2-carboxylic acidethyl ester (Reference Example 23a). This material is dissolved in 50 mLof toluene, then 689 mg, 586 μL (4.47 mmol, 2.0 equiv.) of diethylsulfate and 309 mg (2.24 mmol, 1.0 equiv.) of K₂CO₃ were added. Thereaction was then heated to reflux for 24 hr. At the end of this time,monitoring by LC/MS reveals that the reaction is >than 95% complete. Thereaction is then cooled, 100 mL of ethyl acetate is added and theorganic layer is washed with 0.5N HCl solution, dried over Na₂SO₄,filtered and concentrated. The residues were subjected to flashchromatography, using 40% ethyl acetate in hexane as eluent. Thepurified fractions were concentrated to yield 500 mg (65%) of acolorless solid. Mass Spec.: calc. for [C₁₄H₁₃BrO₅+H]⁺ Theor. m/z=341,343; Obs.=341, 343.

REFERENCE EXAMPLE 23c6-Ethoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic AcidEthyl Ester

Into a 100 mL, 3 neck round bottom flask equipped with a refluxcondenser, magnetic stirrer and nitrogen inlet is added 350 mg (1.03mmol, 1.0 equiv.) of 8-Bromo-6-ethoxy-4-oxo-4H-chromene-2-carboxylicacid ethyl ester (Reference Example 23b), 18.9 mg (0.02 mmol, 0.02equiv.) of tris dibenzylidineacetone dipalladium, 77 mg (0.123 mmol,0.12 equiv.) of racemic 2,2′-bis(diphenylphosphino)-1,1′-binapthyl and 1g of 4 A molecular sieves and 60 mL of dry toluene. To the stirredsuspension is then added 113 mg, 1255 μL, (1.13 mmol, 1.1 equiv.) of1-methylpiperazine, followed by 470 mg (1.44 mmol, 1.4 equiv.) of cesiumcarbonate. The mixture is then heated to 80° C. for 3 days. At the endof this time completion was monitored by LC/MS analysis of an aliquot.When the reaction was determined to be complete it was cooled to roomtemperature then filtered through a plug of diatomaceous earth, withtoluene washing to remove solid by products. Purification by flashchromatography, using a gradient of 5 to 40% methanol in methylenechloride as eluent, yielded 350 mg (75%) of the desired product as ayellow solid. Mass Spec.: calc. for [C₁₉H₂₄N₂O₅+H]⁺ Theor. m/z=361;Obs.=361.

REFERENCE EXAMPLE 23d6-Ethoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic Acid

Into a 125 mL Erlenmeyer equipped with a magnetic stirrer is placed 500mg (1.39 mmol, 1.0 equiv.) of6-Ethoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidethyl ester (Reference Example 23c). This material is dissolved in 30 mLof THF, then 30 mL of methanol are added. To this stirring solution isadded 30 mL of a water containing 64.2 mg (1.53 mmol, 1.1 equiv.) oflithium hydroxide. This mixture is stirred at room temperature for 2 hr.Completion of the reaction is monitored by LC/MS, then 10 mL of 2N HClis added. This mixture is then concentrated, dried and triturated withether to give the product as the hydrochloride salt in quantitativeyield.

Mass Spec.: calc. for [C₁₇H₂₀N₂O₅+H]⁺ Theor. m/z=333; Obs.=333.

REFERENCE EXAMPLE 23e6-Ethoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carbonylChloride

Into a 100 mL round bottom flask equipped with a reflux condenser,nitrogen inlet and magnetic stirrer is placed 250 mg (0.68 mmol, 1.0equiv.) of6-Ethoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride salt (Reference Example 23d) and 20 mL of methylenechloride. To the stirring suspension is then added 129.5 mg, 164 L(1.02mmol, 1.5 equiv.) of oxalyl chloride followed by addition of one drop ofDMF from a 50 microliter syringe to act as catalyst. The mixture isstirred for 2 hours, then concentrated to dryness on a rotary evaporatorunder a nitrogen atmosphere, followed by drying under high vacuum. Thecompleteness of the reaction was ascertained by analysis of an aliquot,which was quenched with a THF solution of methylamine, by LC/MS. Thecrude material was used as obtained in the subsequent amidationreaction.

REFERENCE EXAMPLE 24

Preparation of8-Bromo-6-methoxy-4-(2-trimethylsilanyl-ethoxymethoxy)-quinoline-2-carboxylicAcid Methyl Ester REFERENCE EXAMPLE 24a2-(2-Bromo-4-methoxy-phenylamino)-but-2-enedioic Acid Dimethyl Ester

A solution of 2-bromo-4-methoxy aniline (6.02 g, 29.8 mmol) in 125 mLanhydrous methanol was treated with dimethyl acetylenedicarboxylate(3.70 mL, 30.2 mmol) and the solution was heated at reflux undernitrogen for 8 hours. The reaction mixture was cooled, concentrated, andredissolved in hot methanol. Yellow crystals were obtained by filtration(6.93 g, 68%). A second crop of crystals was obtained from ethanol(0.942 g, 9%). The filtrates were combined and purified by flashchromatography on silica gel using 4:1 hexanes:ethyl acetate to affordan additional 1.63 g (16%) for a total yield of 93%. ¹H NMR (300 MHz,DMSO, d₆) δ 9.60 (s, 1H, NH), 7.26 (d, 1H, J_(m)=2.7 Hz, ArH ₃), 6.93(dd, 1H, J_(o)=8.7, J_(m)=2.7 Hz, ArH ₅), 6.87 (d, 1H, J_(o)=8.7 Hz, ArH₆), 5.34 (s, 1H, C═CH), 3.76 (s, 3H, OCH ₃), 3.68 (s, 3H, CHCO₂CH ₃),3.66 (s, 3H, CNCO₂CH ₃); Mass Spec.: calc. for [C₁₃H₁₄BrNO₅+H]⁺ Theor.m/z=344, 346; Obs. 344, 346.

REFERENCE EXAMPLE 24a8-Bromo-6-methoxy-4-oxo-1,4-dihydro-quinoline-2-carboxylic Acid MethylEster

Dow-Therm (175 mL) was heated to 244° C. and the2-(2-bromo-4-methoxy-phenylamino)-but-2-enedioic acid dimethyl ester(9.50 g, 27.6 mmol) was added as a solid in portions over 7 minuteswhile maintaining a temperature of 230-240° C. The brown reactionmixture was heated at 240-245° C. for 45 minutes and then cooled to roomtemperature. A yellow precipitate formed upon cooling. Approximately 100mL of hexanes were added to the mixture and the solids were isolated byfiltration, washed with additional hexanes, and dried under high vacuumto afford the product as a yellow solid (6.73 g, 78%). ¹H NMR (300 MHz,DMSO, d₆) δ 12.01 (s, 1H, NH), 7.86 (d, 1H, J_(m)=2.7 Hz, ArH ₅), 7.52(s, 1H, C═CH), 7.48 (d, 1H, J_(m)=2.7 Hz, ArH ₇), 3.93 (s, 6H, OCH ₃ andCO₂CH ₃); Mass Spec.: calc. for [C₁₂H₁₀BrNO₄+H]⁺ Theor. m/z=312, 314;Obs. 312, 314.

REFERENCE EXAMPLE 24c8-Bromo-6-methoxy-4-(2-trimethylsilanyl-ethoxymethoxy)-quinoline-2-carboxylicAcid Methyl Ester

A brown solution of8-bromo-6-methoxy-4-oxo-1,4-dihydro-quinoline-2-carboxylic acid methylester (6.73 g, 21.6 mmol) in 100 mL N-methyl pyrolidinone was treatedwith sodium hydride (60% dispersion in oil, 1.028 g, 25.7 mmol). Gasevolution and warming were observed. The reaction was stirred for 10minutes at room temperature under nitrogen. Addition of2-(trimethylsilyl)ethoxymethyl chloride (5.00 mL, 28.3 mmol) resulted ina slightly cloudy, light brown solution. After 2.5 hours at roomtemperature, the reaction mixture was poured into 800 mL water andstirred for 15 minutes. The resulting cream colored precipitate wasisolated by filtration, washed with water, and dried under high vacuumto afford the product as a cream colored solid (9.70 g, quantitativeyield). ¹H NMR (300 MHz, DMSO, d₆) δ 7.976 (d, 1H, J_(m)=2.7 Hz, ArH7),7.79 (s, 1H, C═CH), 7.53 (d, 1H, J_(m)=2.7 Hz, ArH ₅), 5.70 (s, 2H, OCH₂O), 3.99 (s, 6H, OCH ₃ and CO₂CH ₃), 3.88 (t, 2H, J=8.0 Hz, OCH₂CH₂Si), 0.97 (t, 2H, J=8.0 Hz, OCH₂CH ₂Si),), −0.04 (s, 9H, Si(CH ₃)₃;Mass Spec.: calc. for [C₁₈H₂₄BrNO₅Si+H]⁺ Theor. m/z=442, 444; Obs. 442,444.

REFERENCE EXAMPLE 25

Preparation of6-Methoxy-8-(4-methyl-[1,4]diazepan-1-yl)-4-oxo-1,4-dihydro-quinoline-2-carboxylicAcid REFERENCE EXAMPLE 25a6-Methoxy-8-(4-methyl-[1,4]diazepan-1-yl)-4-(2-trimethylsilanyl-ethoxymethoxy)-quinoline-2-carboxylicAcid Methyl Ester

To a clear, light brown solution of2-bromo-6-methoxy-4-(2-trimethylsilanyl-ethoxymethoxy)-quinoline-2-carboxylicacid methyl ester (1.01 g, 2.28 mmol), N-methylhomopiperazine (0.32 mL,2.57 mmol), and 4 Å sieves in 30 mL anhydrous toluene was added Pd₂(dba)₂ (43.8 mg, 0.048 mmol) and BINAP (169.8 mg, 0.27 mmol). Theresulting wine colored solution was treated with cesium carbonate (1.124g, 3.45 mmol). The reaction mixture was heated at reflux under nitrogenfor 21 hours. The pea green reaction mixture was cooled to roomtemperature and concentrated. The crude mixture was purified by flashchromatography on silica gel using a gradient of 95:5 to 40:60 methylenechloride:methanol to afford the desired product as a yellow foam (1.004g, 92%). ¹H NMR (300 MHz, DMSO, d₆) δ 7.67 (s, 1H, ArH ₃), 6.94 (d, 1H,J_(m)=2.4 Hz, ArH ₅), 6.66 (d, 1H, J_(m)=2.4 Hz, ArH ₇), 5.60 (s, 2H,OCH ₂O), 3.94 (s, 3H, CO₂CH ₃), 3.88 (s, 3H, OCH ₃), 3.82 (t, 2H, J=8.0Hz, OCH ₂CH₂Si), 3.75 (bs, 4H, ArNCH ₂CH₂CH₂NCH₃ & ArNCH ₂CH₂N—CH₃),3.45 (bs, 2H, ArNCH₂CH ₂NCH₃), 3.31 (bs, 2H, ArNCH₂CH₂CH ₂NCH₃), 2.83(s, 3H, NCH ₃), 2.28 (bs, 2H ArNCH₂CH ₂ CH₂NCH₃), 0.92 (t, 2H, J=8.0 Hz,OCH₂CH ₂Si), −0.04 (s, 9H, Si(C H ₃)₃; Mass Spec.: calc. for[C₂₄H₃₇N₃O₅Si+H]⁺ Theor. m/z=476; Obs. 476.

REFERENCE EXAMPLE 25b6-Methoxy-8-(4-methyl-[1,4]diazepan-1-yl)-4-oxo-1,4-dihydro-quinoline-2-carboxylicAcid

To a light brown solution of6-methoxy-8-(4-methyl-[1,4]diazepan-1-yl)-4-(2-trimethylsilanyl-ethoxymethoxy)-quinoline-2-carboxylicacid methyl ester (1.00 g, 2.10 mmol) in 18 mL 3:1:1tetrahydrofuran:methanol:water was added lithium hydroxide monohydrate(0.267 g, 6.35 mmol). The reaction mixture was stirred at roomtemperature for 5 hours, acidified to pH 4 with 1 N HCl, and stirred anadditional 20 minutes. The reaction mixture was concentrated and driedunder high vacuum to afford an orange foam. ¹H NMR (300 MHz, DMSO, d₆) δ11.06 (s, 1H, NH), 7.53 (s, 1H, C═CH), 7.00 (d, 1H, J_(m)=2.4 Hz, ArH₅), 6.70 (d, 1H, J_(m)=2.4 Hz, ArH ₇), 4.05-3.99 (m, 2H, ArNCH₂CH₂CH₂NCH₃), 3.87 (s, 3H, OCH ₃), 3.68-3.60 (m, 2H, ArNCH ₂CH₂NCH₃),3.54-3.47 (m, 2H, ArNCH₂CH ₂NCH₃), 3.41-3.26 (m, 2H, ArNCH₂CH₂CH ₂NCH₃),2.82 (d, 3H, J=4.8 Hz, NCH ₃), 2.46-2.41 (m, 1H ArNCH₂CH ₂CH₂NCH₃),2.30-2.25 (m, 1H ArNCH₂CH ₂CH₂NCH₃); Mass Spec.: calc. for[C₁₇H₂₁N₃O₄+H]⁺ Theor. m/z=332; Obs. 332.

REFERENCE EXAMPLE 26

Preparation of6-Fluoro-8-(4-methyl-piperazin-1-yl)-4-oxo-1,4-dihydro-quinoline-2-carboxylicAcid

This compound was prepared via the same procedure described forpreparation of Reference Example 25.

REFERENCE EXAMPLE 27

Preparation of6-Methoxy-8-(4-methyl-[1,4]diazepan-1-yl)-4-(2-trimethylsilanyl-ethoxymethoxy)-quinoline-2-carboxylicAcid (4-morpholin-4-yl-phenyl)-amide REFERENCE EXAMPLE 27a8-Bromo-6-methoxy-4-oxo-1,4-dihydro-quinoline-2-carboxylic Acid

To a light brown solution of8-bromo-6-methoxy-4-(2-trimethylsilanyl-ethoxymethoxy)-quinoline-2-carboxylicacid methyl ester (Reference Example 24c) (4.98 g, 11.3 mmol) in 75 mL3:1:1 tetrahydrofuran:methanol:water was added lithium hydroxidemonohydrate (1.367 g, 32.6 mmol). The reaction was stirred at roomtemperature for 5 hours. The reaction mixture was concentrated and thenpoured into water. The solution was acidified to pH 2 with 1 N HCl andthe resulting solids were isolated by filtration. The solids were thensuspended in methanol and filtered to afford the desired product (2.6732g, 80%). An additional 0.5768 g (17%) of product was obtained from themethanol filtrates. ¹H NMR (300 MHz, DMSO, d₆, TFA Shake) δ 7.86 (d, 1H,J_(m)=2.7 Hz, ArH ₅), 7.55 (d, 1H, J_(m)=2.7 Hz, ArH ₇), 7.32 (s, 1H,C═CH), 3.94 (s, 3H, OCH ₃); Mass Spec.: calc. for [C₁₁H₈BrNO₄+H]⁺ Theor.m/z=298, 300; Obs.=298, 300.

REFERENCE EXAMPLE 27b8-Bromo-6-methoxy-4-oxo-1,4-dihydro-quinoline-2-carboxylic Acid(4-morpholin-4-yl-phenyl)-amide

To a yellow suspension of8-bromo-6-methoxy-4-oxo-1,4-dihydro-quinoline-2-carboxylic acid(Reference Example 27a) (3.446g, 11.56 mmol), TBTU (9.039 g, 28.15mmol), and HOBt (3.757 g, 27.8 mmol) in 100 mL dimethylformamide wasadded 4-morpholinoaniline (2.733 g, 15.3 mmol) and diisopropylethylamine (8.2 mL, 50.2 mmol). The resulting marroon solution was stirred atroom temperature under nitrogen for 16 hours during which time thereaction became greenish brown and formed a large amount of precipitate.The reaction mixture was filtered and the solids washed withdimethylformamide, water, and methanol. Drying under high vacuumafforded the desired product as a yellow solid (3.09 g, 58%). ¹H NMR(300 MHz, DMSO, d₆) δ 12.13 (s, 1H, NH), 10.18 (s, 1H, C(O)NH), 7.90 (d,1H, J_(m)=2.7 Hz, ArH ₅), 7.68 (d, 2H, J_(o)=9.0 Hz, ArH _(2′) & H_(6′)), 7.63 (s, 1H, C═CH), 7.51 (d, 1H, J_(m)=2.7 Hz, ArH ₇), 7.00 (d,2H. J_(o)=9.0 Hz, ArH _(3′) & H _(5′)), 3.94 (s, 3H, OCH ₃), 3.75 (t,4H, J=4.8 Hz, OCH ₂CH₂N), 3.10 (t, 4H, J=4.8 Hz, OCH₂CH ₂N); Mass Spec.:calc. for [C₂₁H₂₀BrN₃O₄+H]⁺ Theor. m/z=458, 460; Obs.=458, 460.

REFERENCE EXAMPLE 27c8-Bromo-6-methoxy-4-(2-trimethylsilanyl-ethoxymethoxy)-quinoline-2-carboxylicacid (4-morpholin-4-yl-phenyl)-amide

A yellow suspension of8-bromo-6-methoxy-4-oxo-1,4-dihydro-quinoline-2-carboxylic acid(4-morpholin-4-yl-phenyl)-amide (Reference Example 27b) (3.092 g, 6.75mmol) in 40 mL N-methylpyrolidinone was treated with sodium hydride (60%dispersion in oil, 0.410 g, 10.24 mmol). Gas evolution and warming wereobserved and the suspension became light brown and almost clear. Thereaction was stirred for 10 minutes at room temperature under nitrogen.Addition of the 2-(trimethylsilyl)ethoxymethyl chloride (1.6 mL, 9.1mmol) resulted in a slightly cloudy, lighter brown solution. After 4.5hours at room temperature, the reaction mixture was poured into 300 mLwater, stirred for 15 minutes and then stored at 0° C. overnight. Thesolids were isolated by filtration, suspended in methanol, filteredagain, and dried under high vacuum to afford the product as a yellowsolid (3.190 g, 80%). ¹H NMR (300 MHz, DMSO, d₆) δ 10.18 (s, 1H,C(O)NH), 7.95 (d, 1H, J_(m)=2.4 Hz, ArH ₇), 7.83 (s, 1H, ArH ₃), 7.69(d, 2H, J_(o)=9.0 Hz, ArH _(2′) & H_(6′)), 7.51 (d, 1H, J_(m)=2.7 Hz,ArH ₅), 7.00 (d, 2H, J_(o)=9.0 Hz, ArH _(3′) & H _(5′)), 5.69 (s, 2H,OCH₂O), 3.95 (s, 3H, OCH ₃), 3.85 (t, 2H, J=8.0 Hz, OCH ₂CH₂Si), 3.75(t, 4H, J=4.7 Hz, OCH ₂CH₂N), 3.10 (t, 4H, J=4.7 Hz, OCH₂CH ₂N), 0.94(t, 2H, J=8.0 Hz, OCH₂CH ₂Si), −0.04 (s, 9H, Si(C H ₃)₃; Mass Spec.:calc. for [C₂₇H₃₄BrN₃O₅Si+H]⁺ Theor. m/z=588, 590; Obs.=588, 590.

REFERENCE EXAMPLE 27d6-Methoxy-8-(4-methyl-[1,4]diazepan-1-yl)-4-(2-trimethylsilanyl-ethoxymethoxy)-quinoline-2-carboxylicAcid (4-morpholin-4-yl-phenyl)-amide

To a yellow-green suspension of8-bromo-6-methoxy-4-(2-trimethylsilanyl-ethoxymethoxy)-quinoline-2-carboxylicacid (Reference Example 27c) (4-morpholin-4-yl-phenyl)-amide (1.155 g,1.96 mmol), N-methyl homopiperazine (0.39 mL, 3.14 mmol), and 4 Å sievesin 30 mL anhydrous toluene was added Pd₂ (dba)₂ (90.0 mg, 0.098 mmol)and BINAP (0.358 g, 0.58 mmol). The resulting reddish brown mixturebecame lighter in color upon treatment with cesium carbonate (2.544 g,7.81 mmol). The reaction mixture was heated at reflux under nitrogen for17 hours. The clear brown solution was cooled to room temperature,concentrated, and then purified by flash chromatography on silica gelusing a slow gradient of 95:5 to 50:50 methylene chloride:methanol toafford the desired product (0.989 g, 81%). ¹H NMR (300 MHz, DMSO, d₆) δ9.88 (s, 1H, NH), 7.73 (s, 1H, ArH ₃), 7.68 (d, 2H, J_(o)=8.9 Hz, ArH_(2′) & H _(6′))7.00 (d, 2H, J_(o)=8.9 Hz, ArH _(3′) & H _(5′)), 6.94(d, 1H, J_(m)=2.7 Hz, ArH ₅), 6.66 (d, 1H, J_(m)=2.7 Hz, ArH ₇), 5.62(s, 2H, OCH₂O), 3.87 (s, 3H, OCH ₃), 3.80(t, 2H, J=8.0 Hz, OCH ₂CH₂Si),3.73 (t, 4H, J=4.7 Hz, OCH ₂CH₂N), 3.63 (t, 2H, J=5.9 Hz, ArNCH₂CH₂CH₂NCH₃), 3.33 (bs, 2H, ArNCH ₂CH₂NCH₃), 3.09 (t, 4H, J=4.7 Hz,OCH₂CH ₂N), 2.97 (bs, 2H, ArNCH₂CH ₂NCH₃), 2.69 (bs, 2H, ArNCH₂CH₂CH₂NCH₃), 2.35 (s, 3H, NCH ₃), 2.09 (bs, 2H ArNCH₂CH ₂CH₂NCH₃), 0.94 (t,2H, J=8.0 Hz, OCH₂CH ₂Si), −0.03 (s, 9H, Si(CH ₃)₃; Mass Spec.: calc.for [C₃₃H₄₇N₅O₅Si+H]⁺ Theor. m/z=622; Obs.=622.

REFERENCE EXAMPLE 28

Preparation of 8-Bromo-4-dimethylamino-6-methoxy-quinoline-2-carboxylicAcid (4-morpholin-4-yl-phenyl)-amide REFERENCE EXAMPLE 28a8-Bromo-4-chloro-6-methoxy-quinoline-2-carboxylic Acid(4-morpholin-4-yl-phenyl)-amide

A suspension of8-bromo-6-methoxy-4-oxo-1,4-dihydro-quinoline-2-carboxylic acid(Reference Example 27b) (1.75 mmol) in 20 mL methylene chloride wastreated with oxalyl chloride (1.5 mL, 17.2 mmol) and catalyticdimethylformamide (3 drops). The reaction mixture bubbled vigorously andbecame clearer. The reaction was heated at reflux for 2 hours, cooled toroom temperature, and concentrated to a pale yellow solid (kept undernitrogen).

To a yellow solution of the acid chloride in 20 mL methylene chloridewas added 4-morpholinoaniline (0.347 g, 1.94 mmol) and diisopropylethylamine (1.0 mL, 6.1 mmol). The solution became orange and gas evolutionwas observed. Within 30 minutes, solids began to precipitate from thesolution. The reaction was stirred at room temperature for 1 hour. Thesolids were isolated by filtration and dried under high vacuum to affordthe desired product (0.406 g, 49%). ¹H NMR (300 MHz, DMSO, d₆) δ 10.15(s, 1H, C(O)NH), 8.33 (s, 1H, ArH ₃), 8.10 (d, 1H, J_(m)=2.7 Hz, ArH ₇),7.70 (d, 2H, J_(o)=9.0 Hz, ArH _(2′) & H _(6′)), 7.56 (d, 1H, J_(m)=2.7Hz, ArH ₅), 7.01 (d, 2H, J_(o)=9.0 Hz, ArH _(3′) & H _(5′)), 4.06 (s,3H, OCH ₃), 3.75 (t, 4H, J=4.8 Hz, OCH ₂CH₂N), 3.11 (t, 4H, J=4.8 Hz,OCH₂CH ₂N); Mass Spec.: calc. for [C₂₁H₁₉BrClN₃O₃+H]⁺ Theor. m/z=476,478; Obs.=476, 478.

REFERENCE EXAMPLE 28b8-Bromo-4-dimethylamino-6-methoxy-quinoline-2-carboxylic Acid(4-morpholin-4-yl-phenyl)-amide

A solution of 8-bromo-4-chloro-6-methoxy-quinoline-2-carboxylic acid(4-morpholin-4-yl-phenyl)-amide (Reference Example 28a) (0.1512 g, 0.317mmol) in 100 mL 2.0 M dimethyl amine in tetrahydrofuran was heated at100° C. in a Parr bomb. The initial pressure was 75-80 psi and thenremained at approximately 60 psi. After 18 hours, the reaction wascooled to room temperature, concentrated and dried to afford the crudeproduct as a brown solid. Purification on silica gel using a gradient of100:0 to 95:5 methylene chloride:methanol afforded the clean product(0.142 g, 92%). ¹H NMR (300 MHz, DMSO, d₆) δ 10.20 (s, 1H, C(O)NH), 7.90(d, 1H, J_(m)=2.7 Hz, ArH ₅), 7.69 (d, 2H, J_(o)=9.0 Hz, ArH _(2′) & H_(6′)), 7.60 (s, 1H, ArH ₃), 7.41 (d, 1H, J_(m)=2.7 Hz, ArH ₇), 7.01 (d,2H, J_(o)=9.0 Hz, ArH _(3′) & H _(5′)), 3.96 (s, 3H, OCH ₃), 3.75 (t,4H, J=4.8 Hz, OCH ₂CH₂N), 3.10 (t, 4H, J=4.8 Hz, OCH₂CH ₂N), 3.08 (s,6H, N(CH ₃)₂); Mass Spec.: calc. for [C₂₁H₁₉BrClN₃O₃+H]⁺ Theor. m/z=485,487; Obs.=485, 487.

REFERENCE EXAMPLE 29

Preparation of6-Fluoro-4-methoxy-8-(4-methyl-piperazin-1-yl)quinoline-2-carboxylicAcid REFERENCE EXAMPLE 29a8-Bromo-6-fluoro-4-methoxy-quinoline-2-carboxylic Acid Methyl Ester

Into a 150 mL 3 neck round bottom flask equipped with a refluxcondenser, magnetic stirrer and nitrogen inlet is placed 2.0 g (6.76mmol, 1.0 equiv.) of8-Bromo-6-fluoro-4-oxo-1,4-dihydro-quinoline-2-carboxylic acid methylester. This material is then dissolved in 50 mL of NMP. Then 300 mg(7.44 mmol, 1.1 equiv.) of a 60% dispersion of sodium hydride in oil iscautiously added portion-wise to the solution at room temperature. Ayellow color then develops, indicating that formation of the anion hasoccurred, with hydrogen evolution. Stirring of the anion solution iscontinued for one hour, then 1.14 g, 500 μl (8.04 mmol, 1.2 equiv.) ofiodomethane is added via syringe. The mixture is allowed to react fortwo hours additional, then is cautiously quenched with 20 mL of water.The solids, which precipitate upon dilution in 1 L of water, arecollected by filtration, then washed with water to give the pure Omethylated material as 2.1 g (98%) of a colorless solid. Mass Spec.:calc. for [C₁₂H₉BrFNO₃+H]⁺ Theor. m/z=314, 316; Obs.=314, 316.

Alternatively, into a 100 mL 3 neck round bottom flask equipped with areflux condenser, nitrogen inlet and magnetic stirrer is placed 350 mg(1.17 mmol, 1.0 equiv.) of8-Bromo-6-fluoro-4-oxo-1,4-dihydro-quinoline-2-carboxylic acid methylester and 242 mg (1.75 mmol, 1.5 equiv.) of K₂CO₃. This material issuspended in 20 mL of DMSO then heated to 70° C. for 1 hr. The anionformation of the anion is apparent when the mixture becomes cloudy. Themixture is allowed to cool to 35° C. then 331 mg, 145 μL (2.33 mmol, 2.0equiv.) of methyl iodide are added and stirring is continued for 2 hr.At the end of this time it is determined if the reaction is complete byLC/MS. Upon completion the mixture is poured into 200 mL of water andthe solids which form are collected by filtration and washed with waterto give 340 mg (93%) of the O-methylated product after drying.

REFERENCE EXAMPLE 29b6-fluoro-4-methoxy-8-(4-methyl-piperazin-1-yl)-quinoline-2-carboxylicAcid Methyl Ester

Into a 250 mL, 3 neck round bottom flask equipped with a refluxcondenser, magnetic stirrer and nitrogen inlet is added 2.1 g (6.68mmol, 1.0 equiv.) of 8-Bromo-6-fluoro-4-methoxy-quinoline-2-carboxylicacid methyl ester (Reference Example 29a) (122 mg, 0.134 mmol, 0.02equiv.) of tris dibenzylidineacetone dipalladium, 499 mg (0.802 mmol,0.12 equiv.) of racemic 2,2′-bis(diphenylphosphino)-1,1′-binapthyl and 1g of 4 A molecular sieves and 80 mL of dry toluene. To the stirredsuspension is then added 736 mg, 815 μL, (7.35 mmol, 1.1 equiv.) of1-methylpiperazine, followed by 3:05 g (9.35 mmol, 1.4 equiv.) of cesiumcarbonate. The mixture is then heated to 80° C. for 36 hr. At the end ofthis time completion was monitored by LC/MS analysis of an aliquot. Whenthe reaction was determined to be complete it was cooled to roomtemperature then filtered through a plug of celite, with toluene washingto remove solid by products. Purification by flash chromatography usinga gradient of 5 to 20% methanol in methylene chloride as eluent yielded2.0 g, (90%) of the desired product. Mass Spec.: calc. for[C₁₇H₂₀FN₃O₃+H]⁺ Theor. m/z=334; Obs.=334.

REFERENCE EXAMPLE 29c6-fluoro-4-methoxy-8-(4-methyl-piperazin-1-yl)-quinoline-2-carboxylicAcid

Into a 125 mL erlenmeyer flask containing 30 mL of THF and 30 mL ofmethanol is placed 2.1 g (6.3 mmol) of6-fluoro-4-methoxy-8-(4-methyl-piperazin-1-yl)-quinoline-2-carboxylicacid methyl ester (Reference Example 29b). To this solution is addedwith stirring 30 mL of water in which is dissolved 291 mg (6.9 mmol, 1.1equiv.) of lithium hydroxide monohydrate. This solution is allowed toreact for 1 hr then is quenched with 10 mL of 2N HCl solution. Thesolution is then filtered and the solids washed with 10 mL of 0.5 N HClsolution. The combined filtrates are then concentrated to give 2.15 g,(95%) of the solid yellow product as the hydrochloride salt. Mass Spec.:calc. for [C₁₆H₁₈FN₃O₃+H]⁺ Theor. m/z=320; Obs.=320.

EXAMPLE 1

8-(4-methyl-1-piperazinyl)-N-[4-(4-morpholinyl)phenyl]-4-oxo-4H-chromene-2-carboxamide

8-(4-methyl-1-piperazinyl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride (Reference Example 1) (400 mg, 1.23 mmol) was suspended inanhydrous N,N-dimethylformamide (20 ml) and triethylamine (0.69 ml, 4.92mmol) was added to give a clear solution. The following were added inorder: 1-hydroxybenzotriazole (HOBt (205 mg, mol)),O-(1H-Benzotriazol-1-yl)-N,N,N′,N′-pentamethylene-uroniumtetrafluoroborate (TBTU (435 mg, 3.1 mmol)) then4-(dimethylamino)pyridine (25 mg). After stirring for 5 min at roomtemperature, 4-(4-morpholinyl)aniline (Reference Example 21) (220 mg,mmol). The reaction stirred overnight at room temperature. The solutionwas concentrated in vacuo, the remains were partitioned betweenchloroform/saturated sodium bicarbonate, extracted (×3) with chloroform,dried (MgSO₄) and concentrated in vacuo to give the crude product.

Chromatography on silica (230-400 mesh ASTM) and eluting ethyl acetatefollowed by 2.5-5% methanol/chloroform gave 190 mg (% yield) of8-(4-methyl-1-piperazinyl)-N-[4-(4-morpholinyl)phenyl]-4-oxo-4H-benzochromene-2-carboxamideas a yellow solid (mp 217-218° decomposition and melt 244-247C). LC/MS(M+1) m/z=449.

EXAMPLE 2

2-{1-[4-(2-Methoxy-phenyl)-piperazin-1-yl]-methanoyl}-8-(4-methyl-piperazin-1-yl)-chromen-4-one

This compound was prepared from8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride (Reference Example 1) and commercially available1-(2-Methoxy-phenyl)-piperazine (Aldrich) via the same procedure used inexample 1, yielding a yellow solid. MS (M+H) m/z=463.

EXAMPLE 3

2-{1-[4-(1-Acetyl-2,3-dihydro-1H-indol-6-yl)-piperazin-1-yl]-methanoyl}-8-(4-methyl-piperazin-1-yl)-chromen-4-one

This compound was prepared from8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride (Reference Example 1) and1-(6-Piperazin-1-yl-2,3-dihydro-indol-1-yl)-ethanone (Reference Example8) as prepared in Example 1, yielding a yellow solid. MS (M+H) m/z=516.

EXAMPLE 4

2-Chloro-5-(4-{1-[8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromen-2-yl]-methanoyl}-piperazin-1-yl)-benzonitrile

This compound was prepared from8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride (Reference Example 1) and 2-chloro-5-piperazin-1-ylbenzonitrile (Reference Example 9) as prepared in Example 1, yielding ayellow solid. MS (M+H) m/z=493.

EXAMPLE 5

2-{1-[4-(4-Methoxy-phenyl)-piperazin-1-yl]-methanoyl}-8-(4-methyl-piperazin-1-yl)-chromen-4-one

This compound was prepared from8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride (Reference Example 1) and commercially available (Aldrich)1-(4-Methoxy-phenyl)-piperazine as prepared in example 1, yielding ayellow solid. MS (M+H) m/z=463.

EXAMPLE 6

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic Acid(5-furan-2-yl-1H-pyrazol-3-yl)-amide

This compound was prepared from8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride (Reference Example 1) and commercially available5-furan-2-yl-1H-pyrazol-3-ylamine (Maybridge) as prepared in example 1,yielding a yellow solid. MS (M+H) m/z=420.

EXAMPLE 7

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic Acid(4-imidazol-1-yl-phenyl)-amide

This compound was prepared from8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride (Reference Example 1) and commercially available4-imidazol-1-yl-phenylamine (Aldrich) as prepared in Example 1, yieldinga yellow solid. MS (M+H)m/z=430.

EXAMPLE 8

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic Acid(4-[1,2,3]thiadiazol-5-yl-phenyl)-amide

This compound was prepared from8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride (Reference Example 1) and4-[1,2,3]thiadiazol-5-yl-phenylamine (Reference Example 10) as preparedin Example 1, yielding a yellow solid. MS (M+H)m/z=448.

EXAMPLE 9

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic Acid4-[1,2,3]thiadiazol-5-yl-benzylamide

This compound was prepared from8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride (Reference Example 1) and commercially available(Maybridge) 4-[1,2,3]thiadiazol-5-yl-benzylamine as prepared in Example1, yielding a yellow solid. MS (M+H) m/z=462.

EXAMPLE 10

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic Acid[4-(4-acetyl-piperazin-1-yl)-phenyl]-amide

This compound was prepared from8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride (Reference Example 1) and1-[4-(4-amino-phenyl)-piperazin-1-yl]-ethanone (Reference Example 11) asprepared in Example 1, yielding a yellow solid. MS (M+H) m/z=499.

EXAMPLE 11

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic Acid[4-(4-methanesulfonyl-piperazin-1-yl)-phenyl]-amide

This compound was prepared from8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride (Reference Example 1) and4-(4-methanesulfonyl-piperazin-1-yl)-phenylamine (Reference Example 12)as prepared in Example 1, yielding a yellow solid. MS (M+H) m/z=526.

EXAMPLE 12

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic Acid(2-methoxy-4-morpholin-4-yl-phenyl)-amide

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride (Reference Example 1) (0.10 g, 0.35 mmol), HOBt (0.10 g,0.7 mmol), TBTU (0.225 g, 0.7 mmol), 4-(dimethylamino) pyridine (0.01 g,catalytic amount), triethylamine (0.15 mL, 1.04 mmol), and commerciallyavailable 2-methoxy-4-morpholin-4-yl-phenylamine (SALOR) (0.08 g, 0.38mmol) were dissolved in dimethylformamide (2.5 mL) and stirred at roomtemperature overnight. Ethyl acetate (150 mL) was added and theresulting mixture was washed with water (3×50 mL), dried (Na₂SO₄),filtered, concentrated under vacuum and triturated with ether to yield ayellow solid (85 mg, 54%). LCMS: m/z=480.3.

EXAMPLE 13

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic Acid(3-chloro-4-morpholin-4-yl-phenyl)-amide

This compound was prepared from8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride (Reference Example 1) and commercially available3-chloro-4-morpholin-4-yl-phenylamine (Maybridge) as prepared in Example12, yielding a yellow solid. (110 mg=73%), LCMS−m/z=483.5.

EXAMPLE 14

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic Acid(4-thiomorpholin-4-yl-phenyl)-amide

This compound was prepared from8-4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride (Reference Example 1) and 4-thiomorpholin-4-yl-phenylamine(Reference Example 13) as prepared in Example 12, yielding a yellowsolid. (55 mg=38%), LCMS−m/z=465.5.

EXAMPLE 15

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic Acid(2,5-diethoxy-4-morpholin-4-yl-phenyl)-amide

This compound was prepared from8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride (Reference Example 1) and commercially available2,5-diethoxy-4-morpholin-4-yl-phenylamine (Aldrich) as prepared inExample 12, yielding a yellow solid. (80 mg=50%), LCMS−m/z=537.6.

EXAMPLE 16

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic Acid(4-cyanomethyl-phenyl)-amide

This compound was prepared from8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride (Reference Example 1) and commercially available(4-amino-phenyl)-acetonitrile (Aldrich) as prepared in Example 12,yielding a yellow solid. (65 mg=54%), LCMS−m/z=403.5.

EXAMPLE 17

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic Acid(1H-indol-5-yl)-amide

This compound was prepared from8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride (Reference Example 1) and commercially available1H-indol-5-ylamine (Aldrich) as prepared in Example 12, yielding ayellow solid. (35 mg 29%), LCMS−mz=401.6.

EXAMPLE 18

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic Acid[4-(1-morpholin-4-yl-methanoyl)-phenyl]-amide

This compound was prepared from8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride (Reference Example 1) and1-(4-amino-phenyl)-1-morpholin-4-yl-methanone (Reference Example 14) asprepared in Example 12, yielding a yellow solid. (21 mg=15%),LCMS−m/z=477.6.

EXAMPLE 19

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic Acid[4-(2,6-dimethyl-morpholin-4-yl)-phenyl]-amide

This compound was prepared from8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride (Reference Example 1) and commercially available4-(2,6-dimethyl-morpholin-4-yl)-phenylamine (Maybridge) as prepared inExample 12, yielding a yellow solid. (60 mg=42%), LCMS−m/z=477.6.

EXAMPLE 20

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic Acid[4-(4-fluoro-phenoxy)-phenyl]-amide

This compound was prepared from8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride (Reference Example 1) and commercially available4-(4-fluoro-phenoxy)-phenylamine (Maybridge) as prepared in Example 12,yielding a yellow solid. (110 mg=77%), LCMS−m/z=475.6.

EXAMPLE 21

8-(4-Methyl-piperazin-1-yl)-2-(6-morpholin-4-yl-benzooxazol-2-yl)-chromen-4-one

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride (Reference Example 1) (0.532 g, 1.85 mmol) was placed in a25 mL 3-neck flask under nitrogen and treated with PPA (6 g). Themixture was then treated with the prepared intermediate4-amino-3-hydroxyphenylmorpholine (0.43 g of ˜85% pure, ˜2 mmol). Themixture was stirred and heated in an oil bath to 205° C. for 3 hours togive a dark liquid. The mixture was cooled to room temperature andtreated with 10 mL of water to give a dark solution. The solution wasslowly neutralized with 1N aqueous sodium hydroxide to pH˜7 as a solidformed. The solid was collected, washed several times with water, airdried, and vacuum dried at room temperature to give 0.65 g of a blacksolid. TLC (10% MeOH in CHCl₃ on SiO₂) showed 2 major components atR_(f)˜0.5 and several lower R_(f) minor components. The solid wastriturated with saturated aqueous sodium bicarbonate at roomtemperature. It was filtered off, washed several times with water, andair dried to give 0.65 g of a dark gray solid. TLC showed the samecomponents seen previously. Mass spectral analysis showed m/e=447 bypositive ion CI and m/e=446 by negative ion CI. The solid was dissolvedin 2% methanol in chloroform and it was chromatographed on a MegabondElute silica gel column (10 g of SiO₂) using 2% methanol in chloroform.The slightly faster R_(f) yellow component was concentrated to give0.0188 g of a yellow solid. CI mass spectral analysis showed m/e=447 asthe base peak by positive ion CI. The solid was recrystallized inmethanol to give 0.0178 g of a yellow solid with a melting point of158.1-158.8° C. Proton NMR (CDCl₃) and CI mass spectral analyses wereconsistent for the desired product (m/z=447 base peak by positive ion CIand m/z=446 base peak by negative ion CI).

EXAMPLE 22

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic Acid(2-hydroxy-4-morpholin-4-yl-phenyl)-amide

8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride (Reference Example 1) (0.3768 g, 1.16 mmol) was placed ina 100 mL 3-neck flask under nitrogen and it was dissolved in 20 mL ofDMF. The solution was treated with triethylamine (0.49 mL, 3.5 mmol)followed by HOBT hydrate (0.36g, 2.3 mmol) followed by TBTU (0.74 g, 2.3mmol) and then followed by DMAP (0.020 g). The mixture was stirred for10 minutes and then it was treated with4-amino-3-hydroxyphenylmorpholine (Reference example 21) (0.228 g, 1.17mmol). The mixture was stirred for 15 minutes and then it was treatedwith triethylamine (0.17 mL, 1.2 mmol). The mixture was stirred at roomtemperature for 42 hours and then it was added to a solution of 50 mL ofsaturated aqueous sodium bicarbonate and 50 mL of water. The mixture wasextracted 4 times with ethyl acetate, dried over magnesium sulfate,filtered, and concentrated to give 0.834 gram of a purple oil. The oilwas dissolved in 2 percent methanol in chloroform and it was placed on asilica gel column (5.5 cm diameter by 10.5 cm long) and eluted with 2percent methanol in chloroform followed by 5 percent methanol inchloroform. The yellow fraction was concentrated to give 0.2031 gram ofan orange-yellow solid. The solid was dissolved in methanol, filteredthrough a medium sintered glass funnel, and concentrated to a few mlvolume as a solid formed. The solid was filtered off, washed withmethanol, and air dried to give 0.1613 gram of a tan solid with MP of248.4-249.6° C. Proton COSY NMR and CI mass spectral analyses wereconsistent for the desired product (m/z=465 by positive ion CI andm/z=463 by negative ion CI).

EXAMPLE 23

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic Acid(5-ethoxy-benzothiazol-2-yl)-amide

This compound was prepared from8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride (Reference Example 1) and commercially available5-ethoxy-benzothiazol-2-ylamine (SALOR) as prepared in Example 12,yielding a yellow solid. (55 mg=39%), LCMS−m/z=465.3.

EXAMPLE 24

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic Acid(4-bromo-phenyl)-amide

This compound was prepared from8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride (Reference Example 1) and commercially available4-bromo-phenylamine (Aldrich) as prepared in Example 12, yielding ayellow solid. (1.0 g=75%), LCMS−m/z=442.4.

EXAMPLE 25

8-(4-Methylpiperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic Acidmethyl-(4-morpholin-4-yl-phenyl)amide

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acid(4-morpholin-4-yl-phenyl)-amide (Example 1) (0.1046 g, 0.2332 mmol) wasplaced in a 10 mL single neck round flask under nitrogen. The solid wasdissolved in 2.8 mL of anhydrous DMF. The yellow solution was stirred atroom temperature and treated with one portion of sodium hydride (0.011 gof 95%, 0.44 mmol). The mixture evolved gas and became a red solution.It was stirred under nitrogen for 20 minutes and then it was treatedwith iodomethane (0.015 mL, 0.033 g, 0.233 mmol). The mixture was sealedand stirred at room temperature for 18 hours.

The reaction mixture was concentrated to remove most of the DMF (35 Cbath @0.5 mm) to give a dark semisolid. It was treated with a few dropsof water followed by 10 mL of ethyl acetate. The mixture was dried overmagnesium sulfate, filtered, and concentrated to give 0.0564 gram of ayellow glass. The glass was triturated with diethyl ether, filtered off,and dried under high vacuum to give 0.0302 g of a tan solid with MP of245.0-246.8 C. Proton NMR and CI mass spectral analyses were consistentfor the desired product (m/z 463 by positive ion CI).

EXAMPLE 26

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic Acid(3-morpholin-4-yl-phenyl)-amide

This compound was prepared from8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride (Reference Example 1) and 3-morpholin-4-yl-phenylamine(Reference Example 18) as prepared in Example 12, yielding a yellowsolid. (120 mg=86%), LCMS−m/z=449.5.

EXAMPLE 27

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic Acid(3-cyano-4-morpholin-4-yl-phenyl)-amide

This compound was prepared from8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride (Reference Example 1) and5-amino-2-morpholin-4-yl-benzonitrile (Reference Example 15) as preparedin Example 12, yielding a yellow solid. (120 mg=82%), LCMS−m/z=474.5.

EXAMPLE 28

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic Acid(3-fluoro-4-morpholin-4-yl-phenyl)-amide

This compound was prepared from8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride (Reference Example 1) and3-fluoro-4-morpholin-4-yl-phenylamine (Reference Example 16) as preparedin example 12, yielding a yellow solid. (120 mg=83%), LCMS−m/z=467.6.

EXAMPLE 29

4-[4-({1-[8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromen-2-yl]-methanoyl}-amino)-phenyl]-piperazine-1-carboxylicAcid Tert-Butyl Ester

This compound was prepared from8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride (Reference Example 1) and4-(4-amino-phenyl)-piperazine-1-carboxylic acid tert-butyl ester(Reference Example 17) as prepared in example 12, yielding a yellowsolid. (260 mg=53%), LCMS−m/z=548.6.

EXAMPLE 30

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic Acid(4-piperazin-1-yl-phenyl)-amide

4-[4-({1-[8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromen-2-yl]-methanoyl}-amino)-phenyl]-piperazine-1-carboxylicacid tert-butyl ester (Example 29) (160 mg, 0.3 mmol) was dissolvedethyl acetate (20 mL) and cooled to 0° C. HCl gas was bubbled in slowlyfor 2 minutes. A solid began to precipitate. Methanol (3-4 mL) was addedto dissolve this solid and HCl gas was bubbled in for another 2 minutes.The mixture was concentrated under reduced pressure and triturated withether and dried under vacuum to yield a tan solid (100 mg, 76%).LCMS/m/z=448.6.

EXAMPLE 31

6-Methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicacid (4-morpholin-4-yl-phenyl)-amide

6-Methoxy-8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicacid hydrochloride (Reference Example 2) (3.0g, 8.5 mmol), TBTU (5.5g,17 mmol), 1-hydroxybenztriazole (2.6g, 17 mmol), 4-dimethylaminopyridine(0.05g, catalytic) and commercially available 4-morpholin-4-yl-aniline(1.66g, 9.3 mmol) were dissolved in dimethylformamide (100 mL).Triethylamine (3.5 mL, 25 mmol was added and this mixture stirred atroom temperature for 17 hours. The reaction mixture was concentratedunder vacuum and the residue was partitioned between chloroform (400 mL)and saturated aqueous sodium bicarbonate (50 mL). The organic layer wasseparated, dried (Na₂SO₄), vacuum-filtered and concentrated undervacuum. The residue was purified by chromatography on silica eluted with2-5% methanol in chloroform and then triturated with ether to yield ayellow powder. (1.6 g=39%) LCMS−m/z=479.5 mp=234-236° C.

EXAMPLE 32

6-Methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicAcid [4-(4-methanesulfonyl-piperazin-1-yl)-phenyl]-amide

This compound was prepared from6-Methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicacid hydrochloride (Reference Example 2) and4-(4-methanesulfonyl-piperazin-1-yl)-phenylamine (Reference Example 12)as prepared in example 1, yielding a yellow solid. GC/MS (EI, M+)m/z=556.

EXAMPLE 33

6-Methoxy-8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicAcid (3-chloro-4-morpholin-4-yl-phenyl)-amide

This compound was prepared from6-Methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicacid hydrochloride (Reference Example 2) and commercially available3-chloro-4-morpholin-4-yl-phenylamine (Maybridge) as prepared in Example12, yielding a yellow solid. (45 mg=31%) LCMS−m/z=513.5.

EXAMPLE 34

6-Methoxy-8-4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic Acid(3-fluoro-4-morpholin-4-yl-phenyl)-amide

This compound was prepared from6-methoxy-8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicacid hydrochloride (Reference Example 2) and3-fluoro-4-morpholin-4-yl-phenylamine (Reference Example 16) as preparedin Example 12, yielding a yellow solid. (55 mg=61%), LCMS−m/z=497.5.

EXAMPLE 35

6-Methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicAcid (2-methoxy-4-morpholin-4-yl-phenyl)-amide

This compound was prepared from6-methoxy-8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicacid hydrochloride (Reference Example 2) and commercially available2-methoxy-4-morpholin-4-yl-phenylamine (SALOR) as prepared in Example12, yielding a yellow solid. (55 mg=38%), LCMS−m/z=510.5.

EXAMPLE 36

6-Methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicAcid (4-thiomorpholin-4-yl-phenyl)-amide

This compound was prepared from6-methoxy-8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicacid hydrochloride (Reference Example 2) and4-thiomorpholin-4-yl-phenylamine (Reference Example 13) as prepared inExample 12, yielding a yellow solid. (99 mg=71%), LCMS−m/z=495.5.

EXAMPLE 37

6-Methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicAcid [4-(2,6-dimethyl-morpholin-4-yl)-phenyl]-amide

This compound was prepared from6-methoxy-8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicacid hydrochloride (Reference Example 2) and commercially available4-(2,6-dimethyl-morpholin-4-yl)-phenylamine (Maybridge) as prepared inExample 12, yielding a yellow solid. (70 mg=49%), LCMS−m/z=507.5.

EXAMPLE 38

6-Methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicAcid (3-morpholin-4-yl-phenyl)-amide

This compound was prepared from6-methoxy-8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicacid hydrochloride (Reference Example 2) and3-morpholin-4-yl-phenylamine (Reference Example 18) as prepared inExample 12, yielding a yellow solid. (80 mg=60%), LCMS−m/z=479.5.

EXAMPLE 39

6-Methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicAcid {4-[4-(2-hydroxy-ethyl)-piperazin-1-yl]-phenyl}-amide

This compound was prepared from6-methoxy-8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicacid hydrochloride (Reference Example 2) and2-[4-(4-amino-phenyl)-piperazin-1-yl]-ethanol (Reference Example 19) asprepared in Example 12, yielding a yellow solid. (80 mg=60%).mp=211.5-212.2 (dec.), MS−base peak at m/z=492 by positive ion andm/z=490 by negative ion CI.

EXAMPLE 40

6-Methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicAcid [4-(1-morpholin-4-yl-methanoyl)-phenyl]-amide

This compound was prepared from6-methoxy-8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicacid hydrochloride (Reference Example 2) and1-(4-amino-phenyl)-1-morpholin-4-yl-methanone (Reference Example 14) asprepared in Example 12, yielding a yellow solid. (170 mg=80%),LCMS−m/z=507.5.

EXAMPLE 41

6-Methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicAcid (3-cyano-4-morpholin-4-yl-phenyl)-amide

This compound was prepared from6-methoxy-8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicacid hydrochloride (Reference Example 2) and5-amino-2-morpholin-4-yl-benzonitrile (Reference Example 15) as preparedin Example 12, yielding a yellow solid. (120 mg=57%), LCMS−m/z=504.5.

EXAMPLE 42

4-[4-({1-[6-Methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromen-2-yl]-methanoyl}-amino)-phenyl]-piperazine-1-carboxylicAcid Tert-Butyl Ester

The 6-methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicacid hydrochloride (Reference Example 2) (1.04 g, 2.93 mmol) was placedin a 250 ml 3-neck flask under nitrogen and it was dissolved in 50 ml ofDMF. The solution was treated with triethylamine (1.22 mL, 8.79 mmol)followed by HOBT hydrate (0.90 g, 5.9 mmol) followed by TBTU (1.88 g,5.9 mmol) and then followed by DMAP (0.056 g, 0.46 mmol). The mixturewas stirred for 10 minutes and then it was treated with4-4-Amino-phenyl)-piperazine-1-carboxylic acid tert-butyl ester(Reference Example 17) (0.81 g, 2.9 mmol). The mixture was stirred for15 minutes and then it was treated with triethylamine (0.41 mL, 2.9mmol). The mixture was stirred at room temperature for 18 hours and thenit was concentrated (1 mm Hg pressure, 45 C bath) to give a dark liquid.The concentrate was treated with 80 mL of saturated aqueous sodiumbicarbonate and extracted with ethyl acetate forming a suspended yellowsolid in the organic layer. The solid was filtered off, washed withdiethyl ether, washed with water, and vacuum dried (0.1 mm Hg pressure@25 C) to give 0.36 gram of a yellow solid, M.P.=232.3-232.8 C.

Proton NMR and CI mass spectral analyses were consistent for the desiredproduct (m/e=578 by positive ion CI and m/e=576 by negative ion CI).

The aqueous layer was extracted twice with ethyl acetate, dried overmagnesium sulfate, filtered, and concentrated to give 1.35 gram of adark semisolid. It was triturated with diethyl ether and allowed tostand at room temperature as a solid formed. The solid was filtered off,washed with diethyl ether, and vacuum dried at room temperature to give0.4816 gram of a yellow solid. CI mass spectral analyses was consistentfor the desired product (M/Z=578 BY positive ion CI AND M/Z=576 bynegative ion CI).

EXAMPLE 43

6-Methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicacid (4-piperazin-1-yl-phenyl)-amide

The4-[4-({1-[6-Methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromen-2-yl]-methanoyl}-amino)-phenyl]-piperazine-1-carboxylicacid tert-butyl ester (Example 42) (0.792 gram, 1.37 mmol) was placed ina 50 ml round flask under nitrogen and it was dissolved in 15 ml ofmethylene chloride. The solution was treated with 15 ml oftrifluoroacetic acid (195 mmol) to give a dark solution and it wasstirred at room temperature for 18 hours. It was concentrated to give abrown foam. The foam was treated with 30 ml of saturated aqueous sodiumbicarbonate and it was stirred at room temperature as a yellow solidformed. The solid was filtered off, washed several times with water, airdried and dried under high vacuum (0.1 mm Hg pressure) to give 0.493gram of a yellow solid, M.P.=203.6-204.7 C.

Proton NMR and CI mass spectral analyses were consistent for the desiredproduct (m/z=478 by positive ion CI and m/z=476 by negative ion CI).

EXAMPLE 44-54

The following examples were prepared in parallel by acylation of6-methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicacid (4-piperazin-1-yl-phenyl), amide (Example 43) in an Argonaut Questsynthesizer.

The piperazine side chain was derivatized in parallel fashion usingeleven different commercially available acylating and sulfonatingreagents. The resins used were Argonaut Tech polystyrene amine resins.Each 5 ml Quest tube was charged with 0.010 gram (0.021 mmol) of thestarting N—H piperazine and 3 ml of methylene chloride followed by 4equivalents (0.08 mmol) of PS-DIEA resin (diisopropylbenzylamine PSresin) to scavenge HCl. Each tube was then treated with an acylchloride, sulfonyl chloride, or isocyanate (2 equivalents of each)followed by a little more methylene chloride. The tubes were sealedunder nitrogen, and stirred for 3 hours at room temperature. Themixtures were then opened and treated with about 4 equivalents (0.08mmol) of PS-trisamine resin (primary amine PS resin) to scavenge anyexcess acylating or sulfonating reagent. The mixtures were sealed andstirred for 1.5 hours and then filtered directly into vials andconcentrated to give the products. The products were characterized byHPLC mass spectral analysis and were found to be greater than 90% pureby HPLC. The compounds were submitted to the 5-HT1b binding assay fordetermination of 5-HT receptor binding affinities and selectivities.

EXAMPLE 44

6-Methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicAcid [4-(4-propionyl-piperazin-1-yl)-phenyl]-amide

This compound was prepared from6-methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicacid (4-piperazin-1-yl-phenyl)-amide (Example 43) and commerciallyavailable propionyl chloride (Aldrich) via the parallel synthesisdescribed above. MS−base peak at m/z=534 by positive ion CI.

EXAMPLE 45

6-Methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicAcid [4-(4-ethane sulfonyl-piperazin-1-yl)-phenyl]-amide. MS−Base Peakat m/z=570 by Positive Ion CI

This compound was prepared from6-methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicacid (4-piperazin-1-yl-phenyl)-amide (Example 43) and commerciallyavailable ethanesulfonyl chloride (Aldrich) via the parallel synthesisdescribed above.

EXAMPLE 46

6-Methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicAcid [4-(4-dimethyl sulfamoyl-piperazin-1-yl)-phenyl]-amide

This compound was prepared from6-methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicacid (4-piperazin-1-yl-phenyl)-amide (Example 43) and commerciallyavailable dimethylsulfamoyl chloride (Aldrich) via the parallelsynthesis described above. MS−base peak at m/z=585 by positive ion CI.

EXAMPLE 47

4-[4-({1-[6-Methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromen-2-yl]-methanoyl}-amino)-phenyl]-piperazine-1-carboxylicAcid Dimethylamide

This compound was prepared from6-methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicacid (4-piperazin-1-yl-phenyl)-amide (Example 43) and commerciallyavailable dimethylcarbamyl chloride (Aldrich) via the parallel synthesisdescribed above. MS−base peak at m/z=549 by positive ion CI.

EXAMPLE 48

4-[4-({1-[6-Methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromen-2-yl]-methanoyl}-amino)-phenyl]-piperazine-1-carboxylicAcid Ethylamide

This compound was prepared from6-methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicacid (4-piperazin-1-yl-phenyl)-amide (Example 43) and commerciallyavailable ethyl isocyanate (Aldrich) via the parallel synthesisdescribed above. MS−base peak at m/z=549 by positive ion CI.

EXAMPLE 49

4-[4-({1-[6-Methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromen-2-yl]-methanoyl}-amino)-phenyl]-piperazine-1-carboxylicAcid Cyclohexylamide

This compound was prepared from6-methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicacid (4-piperazin-1-yl-phenyl)-amide (Example 43) and commerciallyavailable cyclohexyl isocyanate (Aldrich) via the parallel synthesisdescribed above. MS−base peak at m/z=603 by positive ion CI.

EXAMPLE 50

4-[4-({1-[6-Methoxy-8-4-methyl-piperazin-1-yl)-4-oxo-4H-chromen-2-yl]-methanoyl}-amino)-phenyl]-piperazine-1-carboxylicAcid Cyclopentylamide

This compound was prepared from6-methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicacid (4-piperazin-1-yl-phenyl)-amide (Example 43) and commerciallyavailable cyclopentanecarbonyl chloride (Aldrich) via the parallelsynthesis described above. MS−base peak at m/z=574 by positive ion CI.

EXAMPLE 51

6-Methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicAcid {4-[4-(1-pyrrolidin-1-yl-methanoyl)-piperazin-1-yl]-phenyl}-amide

This compound was prepared from6-methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicacid (4-piperazin-1-yl-phenyl)-amide (Example 43) and commerciallyavailable 1-pyrrolidinecarbonyl chloride (Aldrich) via the parallelsynthesis described above. MS−base peak at m/z=575 by positive ion CI.

EXAMPLE 52

6-Methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicAcid {4-[4-(propane-2-sulfonyl)-piperazin-1-yl]-phenyl}-amide

This compound was prepared from6-methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicacid (4-piperazin-1-yl-phenyl)-amide (Example 43) and commerciallyavailable isopropylsulfonylonyl chloride (Aldrich) via the parallelsynthesis described above. MS−base peak at m/z=584 by positive ion CI.

EXAMPLE 53

6-Methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicAcid {4-[4-(2-methyl-propanoyl)-piperazin-1-yl]-phenyl}-amide

This compound was prepared from6-methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicacid (4-piperazin-1-yl-phenyl)-amide (Example 43) and commerciallyavailable isobutyryl chloride (Aldrich) via the parallel synthesisdescribed above. MS−base peak at m/z=548 by positive ion CI.

EXAMPLE 54

6-Methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicacid {4-[4-(1-morpholin-4-yl-methanoyl)-piperazin-1-yl]-phenyl}-amide

This compound was prepared from6-methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicacid (4-piperazin-1-yl-phenyl)-amide (Example 43) and commerciallyavailable morpholine-4-carbonyl chloride (Aldrich) via the parallelsynthesis described above. MS−base peak at m/z=591 by positive ion CI.

EXAMPLE 55

6-Fluoro-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acid(4-morpholin-4-yl-phenyl)-amide

This compound was prepared from6-Fluoro-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride (Reference Example 3) and 4-morpholin-4-yl-phenylamine(Reference Example 20) as prepared in Example 1, yielding a yellowsolid. MS (M+I) m/z=467.

EXAMPLE 56

6-Fluoro-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic Acid[4-(4-methanesulfonyl-piperazin-1-yl)-phenyl]-amide

This compound was prepared from6-Fluoro-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride (Reference Example 3) and4-(4-methanesulfonyl-piperazin-1-yl)-phenylamine (Reference Example 12)as prepared in Example 1, yielding a yellow solid. MS (M+H) m/z=544.

EXAMPLE 57

6-Fluoro-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acid[4-(4-acetyl-piperazin-1-yl)-phenyl]-amide

This compound was prepared from6-Fluoro-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride (Reference Example 3) and1-[4-(4-amino-phenyl)-piperazin-1-yl]-ethanone (Reference Example 11) asprepared in Example 1, yielding a yellow solid. MS (M+H) m/z=508.

EXAMPLE 58

6-Fluoro-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic Acid(3-chloro-4-morpholin-4-yl-phenyl)-amide

6-Fluoro-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride (Reference Example 3) (150 mg, 0.43 mmol),1-hydroxybenzotriazole (140 mg, 0.9 mmol),O-(1H-Benzotriazol-1-yl-N,N,N′,N′-pentamethylene-uroniumtetrafluoroborate (290 mg, 0.9 mmol), 4-(dimethylamino)pyridine (10 mg,catalytic), triethylamine (0.2 mL, 1.5 mmol), and commercially available3-chloro-4-morpholin-4-yl-phenylamine (Maybridge) were dissolved indimethylformamide (2.5 mL) and stirred at room temperature overnight. At17 h, water (20 mL) was added and the resulting mixture was stirred for15-30 min. The mixture was vacuum-filtered and the residue washed withwater and air-dried to yield a yellow powder (220 mg=quantitativeyield). LC/MS−m/z=501.5.

EXAMPLE 59

6-Fluoro-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acid(3-fluoro-4-morpholin-4-yl-phenyl)-amide

This compound was prepared from6-Fluoro-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride (Reference Example 3) and3-fluoro-4-morpholin-4-yl-phenylamine (Reference Example 16) as preparedin Example 58, yielding a yellow solid (210 mg=99%). LC/MS−m/z=485.5.

EXAMPLE 60

6-Fluoro-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acid(3-cyano-4-morpholin-4-yl-phenyl)-amide

This compound was prepared from6-Fluoro-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride (Reference Example 3) and5-amino-2-morpholin-4-yl-benzonitrile (Reference Example 15) as preparedin Example 58, yielding a yellow solid (210 mg=99%). LC/MS−m/z=492.5.

EXAMPLE 61

6-Fluoro-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acid[4-(1-morpholin-4-yl-methanoyl)-phenyl]-amide

This compound was prepared from6-Fluoro-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride (Reference Example 3) and1-(4-amino-phenyl)-1-morpholin-4-yl-methanone (Reference Example 14) asprepared in Example 58, yielding a yellow solid (220 mg=quantitativeyield). LC/MS−m/z=495.5.

EXAMPLE 62

6-Methyl-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic Acid(4-morpholin-4-1-phenyl)-amide

This compound was prepared from6-Methyl-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride (Reference Example 4) and 4-morpholin-4-yl-phenylamine(Reference Example 20) as prepared in Example 1, yielding a yellowsolid. LCMS−m/z=463.6.

EXAMPLE 63

6-Methyl-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic Acid[4-(1-morpholin-4-yl-methanoyl)-phenyl]-amide

This compound was prepared from6-Methyl-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride Reference Example 4) and1-(4-amino-phenyl)-1-morpholin-4-yl-methanone (Reference Example 14) asprepared in Example 1, yielding a yellow solid. LCMS−m/z=491.6.

EXAMPLE 64

6-Methyl-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic Acid(3-fluoro-4-morpholin-4-yl-phenyl)-amide

This compound was prepared from6-Methyl-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride (Reference Example 4) and3-fluoro-4-morpholin-4-yl-phenylamine(Reference Example 16) as preparedin Example 1, yielding a yellow solid. LCMS−m/z=504.5.

EXAMPLE 65

6-Chloro-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic Acid(4-morpholin-4-yl-phenyl)-amide

This compound was prepared from6-chloro-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride (Reference Example 5) and 4-morpholin-4-yl-phenylamine(Reference Example 20) as prepared in Example 1, yielding a yellowsolid. LCMS−m/z=483.3.

EXAMPLE 66

5-Methyl-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic Acid(4-morpholin-4-yl-phenyl)-amide

This compound was prepared from5-methyl-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride (Reference Example 6) and 4-morpholin-4-yl-phenylamine(Reference Example 20) as prepared in Example 1, yielding a yellow solid(116 mg 84%) LCMS−m/z=463.5.

EXAMPLE 67

5-Methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicAcid (4-morpholin-4-yl-phenyl)-amide

This compound was prepared from5-methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicacid hydrochloride (Reference Example 7) and4-morpholin-4-yl-phenylamine (Reference example 20) as prepared inExample 1, yielding a yellow solid (149 mg=50%) LCMS−m/z=479.4.

The following additional examples incorporate 4-substitutedpiperazine-1-yl-phenyl amides similar in structure to Examples 44-54.

EXAMPLE 68

6-Methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicacid {4-[4-(3-hydroxy-propanoyl)-piperazin-1-yl]-phenyl}-amide

6-Methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicacid (4-piperazin-1-yl-phenyl)-amide (Example 43) (1.5 gram, 2.12 mmol)was placed in a 100 mL flask with 50 mL of CH₂Cl₂. This suspension wastreated with triethylamine (4 equivalents, 1.2 mL, 8.5 mmol) andβ-propionylactone (0.2 mL, 3.2 mmol) and the reaction stirred at roomtemperature for 2 hours, then heated to 50° C. for 2 hours. Then 0.8 mLmore of b-propionylactone was added and the reaction heated for 4 hoursmore. The reaction was allowed to cool to room temperature and thenconcentrated (1 mm Hg pressure). The concentrate was treated withsaturated aqueous sodium bicarbonate and the resulting solid collectedby vacuum filtration. The residue was purified by chromatography onsilica eluting with 2% methanol in chloroform, then concentrated (1 mmHg pressure). Then triturated with either to yield a yellow powder withwas dried under high vacuum for 48 h at 50° C. (100 mg) LCMS−m/z 550,mp=195-197° C.

EXAMPLE 69

4-[4-({1-[6-Fluoro-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromen-2-yl]-methanoyl}-amino)-phenyl]-piperazine-1-carboxylicAcid Tert-Butyl Ester

This compound was prepared from6-Fluoro-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride (Reference Example 3) and 4{4-Amino-phenyl)-piperazine-1-carboxylic acid tert-butyl ester(Reference Example 17) according to the method of Example 42 to yield(1.65 grams, 64%) of a yellow powder LCMS−m/z=556; mp=219-220° C.

EXAMPLE 70

4-[4-({1-[6-Fluoro-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicAcid (4-piperazin-1-yl-phenyl)-amide

This compound was prepared from4-[4-({-[6-Fluoro-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromen-2-yl]-methanoyl}-amino)-phenyl]-piperazine-1-carboxylicacid tert-butyl ester, as prepared in Example 69, using the method ofExample 43 to yield a yellow solid LCMS−m/z=466.

EXAMPLE 71

6-Fluoro-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic Acid[4-(4-ethane sulfonyl-piperazin-1-yl)-phenyl]-amide

4-[4-({1-[6-Fluoro-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicacid (4-piperazin-1-yl-phenyl)-amide ditrifluoroacetate (the free acidof which was prepared as in Example 70) (4.0 grams, 5.77 mmol) wasplaced in a flask with 50 mL of CH₂Cl₂ and triethylamine (3.2 mL and 23mmol) and ethylsulfonyl chloride was added (0.6 mL, 6.35 mmol)portionwise (0.1 mL at a time) over 15 minutes and allowed to stir atroom temperature for 20 hours. The reaction was concentrated (1 mm Hgpressure) and then saturated aqueous sodium bicarbonate was added andextracted with CHCl₃. The organic fractions were combined, washed withsaturated sodium chloride, dried (MgSO₄) concentrated (1 mm Hg pressure)to give a yellow solid which was recrystallized from methanol to give1.33 grams of product LCMS−m/z=558, mp=233-234° C.

EXAMPLE 72

6-Fluoro-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acid[4-(4-propionyl-piperazin-1-yl)-phenyl]-amide

4-[4-({1-[6-Fluoro-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicacid (4-piperazin-1-yl-phenyl)-amide ditrifluoroacetate (the free acidof which was prepared as in Example 70) (0.69 grams, 1,00 mmol) wasplaced in a flask with 25 mL of CH₂Cl₂ and triethylamine (0.56 mL and 4mmol) and propionyl chloride was added (0.95 mL, 1.1 mmol) and thereaction allowed to stir at room temperature for 20 hours. The residuewas purified by chromatography on silica eluting with 2% methanol inchloroform, then concentrated (1 mm Hg pressure). The residue wastriturated with either then digested with CHCl₃ and the CHCl₃concentrated to yield a yellow powder which was dried under high vacuumfor 48 h at 45° C. (260 mg) LCMS−m/z=522.

EXAMPLE 73

6-Fluoro-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acid{4-[4-(3-hydroxy-propanoyl)-piperazin-1-yl]-phenyl}-amide

This compound was prepared from6-Fluoro-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acid(4-piperazin-1-yl-phenyl)-amide and β-propionylactone using the methoddescribed above in Example 68 to yield 65 mg of a yellow powderLCMS−m/z=538, mp=195-199° C.

The following exemplifies a substituted chromene-2-“reverse amide” (orsubstituted chromene-2-yl-benzamide).

EXAMPLE 74

N-[8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromen-2-yl]-4-morpholin-4-yl-benzamide

8-(4-Methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acidhydrochloride Reference Example 1 (227 mg, 0.69 mmol), triethylamine (2equivalents, 1.389 mmol, 0.193 mL) and diphenylphosphoryl azide (0.69mmol, 0.15 mL) were stirred in toluene (10 mL) at 65° C. for 30 minutes.The reaction was allowed to cool to 22° C. and 4-morpholinobenzonoicacid (0.7 mmol, 145 mg), more triethylamine (0.051 mL, 0.7 mmol), andCH₃CN (5 mL) were added and the reaction heated to reflux for 1 hour.The reaction was concentrated (1 mm Hg pressure) the residue waspartitioned between 1N methanesufonic acid and ether. The acid layer wasthen basified with solid K₂CO₃ and the product extracted in to CHCl₃.The organic layer was dried (MgSO₄) and concentrated under reducedpressure to leave a yellow solid which was further purified with silicachromatography using CHCl₃ to 4% CH₃OH in CHCl₃. Concentration of thefractions containing product yielded 13 mg of product LC/MS−m/z=449.

Enantiomers of 8-(4-Methyl-piperazin-1-yl)-chroman-2-carboxylic acid(4-morpholin-4-yl-phenyl)-amide.

EXAMPLE 75

racemic-8-(4-Methyl-piperazin-1-yl)-chroman-2-carboxylic Acid(4-morpholin-4-yl-phenyl)-amide

racemic-8-(4-Methyl-1-piperazin-1-yl)-chroman-2-carboxylic acidhydrochloride (Example 75a) (1.04 mmol) was dissolved in anhydrousN,N-dimethylformamide (40 ml) and the following were added in order:HOBt (0.17 g, 1.14 mmol), TBTU (0.37 g, 1.14 mmol) then triethylamine(0.6 ml, 4.2 mmol). After stirring for 5 min at room temperature,4-(4-morpholinyl)aniline (reference example 20) (0.185 g, 1.14 mmol) wasadded and the reaction stirred overnight at room temperature.

The solution was concentrated in vacuo, the remains were partitionedbetween chloroform/saturated sodium bicarbonate, extracted (×3) withchloroform, dried (MgSO₄) and concentrated in vacuo to give the crudeproduct.

The crude product was chromatographed on a Waters Delta Prep 4000 using1 PrepPak cartridge (Porasil 37-55 μm 125 Å) eluting with 2.5%methanol/chloroform. The product was collected to give a yellow oil.Ethyl acetate was added to the oil. The solution was refluxed thencooled the yellow solid was filtered to give 55 mg (12% yield) ofracemic-8-(4-methyl-piperazin-1-yl)-chroman-2-carboxylic acid(4-morpholin-4-yl-phenyl)-amide (mp 215-216° C.). The mother liquorcontained 76 mg that was used in the chiral separation described below.LC/MS (M+1) m/z=437.

EXAMPLE 75a racemic-8-(4-Methyl-1-piperazin-1-yl)-chroman-2-carboxylicAcid Hydrochloride

Ethyl 8-(4-methyl-1-piperazin-1-yl)-4-oxo-4H-chromen-2-carboxylate(Reference Example 1) (0.74 g, 2.3 mmol) was dissolved in glacial aceticacid (50 ml) and 10% palladium on carbon (80 mg) was added. The mixturewas hydrogenated on a Paar apparatus (50 psi) at 70° C. for 3 h. Then,concentrated HCl and 10% palladium on carbon (100 mg) were added and themixture was again subjected to hydrogenation (50 psi) at 70° C. for 1 h.The reaction was allowed to cool, the catalyst was filtered and thesolution was concentrated in vacuo. Toluene was repeatedly added and thesolution concentrated to giveracemic-8-(4-Methyl-1-piperazin-1-yl)-chroman-2-carboxylic acidhydrochloride as a foam that was used without further purification inthe next reaction. LC/MS (M+1) m/z=277.

EXAMPLE 76

(+)-8-(4-Methyl-piperazin-1-yl)-chroman-2-carboxylic Acid(4-morpholin-4-yl-phenyl)-amide

The enantiomers ofracemic-8-(4-Methyl-piperazin-1-yl)-chroman-2-carboxylic acid(4-morpholin-4-yl-phenyl)-amide (Example 75) (0.52 g, 1.19 mmol) wereseparated by the use of a chiral column (ChiralPak AD, 5 cm×50 cm, 20μ).The faster (+) isomer (example 76) was eluted with 45%isopropanol/hexane and the slower (−)isomer (example 77) was eluted with75% isopropanol/hexane.

The faster (+) isomer (example 76) was obtained as a white solid (250mg, mp 206-207° C., α_(D)+92.66 in dichloromethane). LC/MS (M+1)m/z=437.

EXAMPLE 77

(−)-8-(4-Methyl-piperazin-1-yl)-chroman-2-carboxylic acid(4-morpholin-4-yl-phenyl)-amide

The enantiomers ofracemic-8-(4-Methyl-piperazin-1-yl)-chroman-2-carboxylic acid(4-morpholin-4-yl-phenyl)-amide (Example 75) (0.52 g, 1.19 mmol) wereseparated by the use of a chiral column (ChiralPak AD, 5 cm×50 cm, 20μ).The faster (+) isomer (example 76) was eluted with 45%isopropanol/hexane and the slower (−)isomer (example 77) was eluted with75% isopropanol/hexane.

The slower (−) isomer (example 77) was obtained as obtained as a lightpurple solid (260 mg, mp 205.5-207° C., α_(D)−91.08 in dichloromethane).LC/MS (M+1) m/z=437.

Enantiomers of 8-(4-methyl-piperazin-1-yl)-4-oxo-chroman-2-carboxylicacid (4-morpholin-4-yl-phenyl)-amide.

EXAMPLE 78

racemic-8-(4-methyl-piperazin-1-yl)-4-oxo-chroman-2-carboxylic Acid(4-morpholin-4-yl-phenyl)-amide

Racemic-8-(4-methyl-1-piperazin-1-yl)-4-oxo-chroman-2-carboxylic acidhydrochloride (Example 78a) (1.04 mmol) was dissolved in anhydrousN,N-dimethylformamide (40 ml) and the following were added in order:HOBt (0.17 g, 1.14 mmol), TBTU (0.37 g, 1.14 mmol) then triethylamine(0.6 ml, 4.2 mmol). After stirring for 5 min at room temperature,4-(4-morpholinyl)aniline (reference example 20) (0.185 g, 1.14 mmol) wasadded and the reaction stirred overnight at room temperature.

The solution was concentrated in vacuo, the remains were partitionedbetween chloroform/saturated sodium bicarbonate, extracted (×3) withchloroform, dried (MgSO₄) and concentrated in vacuo to give the crudeproduct.

The crude product was chromatographed on a Waters Delta Prep 4000 using1 PrepPak cartridge (Porasil 37-55 μm 125 Å) eluting with 2.5%methanol/chloroform. The product was collected to give a yellow oil.Ethyl acetate was added to the oil. The solution was refluxed thencooled the yellow solid was filtered to give 55 mg (12% yield) ofracemic-8-(4-methyl-piperazin-1-yl)-4-oxo-chroman-2-carboxylic acid(4-morpholin-4-yl-phenyl)-amide (mp 215-216° C.). The mother liquorcontained 76 mg that was used in the chiral separation described below.LC/MS (M+) m/z=451.

EXAMPLE 78aracemic-8-(4-Methyl-1-piperazin-1-yl)-4-oxo-chroman-2-carboxylic AcidHydrochloride

racemic-Ethyl-8-(4-methyl-1-piperazinyl)-4-oxo-chroman-2-carboxylate(Example 78b) (0.33 g, 1.04 mmol) was dissolved in 6 M HCl (20 ml) andheated to 100° C. for 1.5 h. The reaction was allowed to cool. Thesolution was concentrated in vacuo and anhydrous toluene was added (×3)and the solution was again concentrated in vacuo to giveracemic-8-(4-Methyl-1-piperazin-1-yl)-4-oxo-chroman-2-carboxylic acidhydrochloride as a yellow foam (0.44 g, quantitative yield) that wasused as is in the next reaction. LC/MS (M+1) m/z=291.

EXAMPLE 78bracemic-Ethyl-8-(4-methyl-1-piperazin-1-yl)-4-oxo-chroman-2-carboxylate

Racemic-Ethyl-8-(4-methyl-1-piperazin-1-yl)-4-hydroxy-chroman-2-carboxylate(Example 78c) (0.43 g, 1.3 mmol) was dissolve in anhydrousdichloromethane (35 ml) and manganese dioxide (1.2 g, 13 mmol) wasadded. The reaction stirred at room temperature overnight.

The reaction was filtered through diatomaceous earth and the solvent wasremoved in vacuo to giveracemic-Ethyl-8-(4-methyl-1-piperazin-1-yl)-4-oxo-chroman-2-carboxylateas a white solid (0.37 g, 86% yield) that was used as is in the nextreaction. GC/MS (EI, M+) m/z=318.

EXAMPLE 78cracemic-Ethyl-8-(4-methyl-1-piperazin-1-yl)-4-hydroxy-chroman-2-carboxylate

Ethyl 8-(4-methyl-1-piperazin-1-yl)-4-oxo-4H-chroman-2-carboxylate(reference example 1) (0.48 g, 1.5 mmol) was dissolved in glacial aceticacid (50 ml) and 10% palladium on carbon (100 mg) was added. The mixturewas hydrogenated on a Paar apparatus (50 psi) at 70° C. for 3 h.

The reaction was allowed to cool, the catalyst was filtered and thesolution was concentrated in vacuo. Ethyl acetate/saturated sodiumbicarbonate was added to the remains and the mixture was extracted (×3)with ethyl acetate, dried (MgSO₄) and stripped to giveracemic-Ethyl-8-(4-methyl-1-piperazin-1-yl)-4-hydroxy-chroman-2-carboxylate(0.43 g, 90% yield) as a yellow oil. GC/MS (EI, M+) m/z=320.

EXAMPLE 79

8-(4-Methyl-piperazin-1-yl)-4-oxo-chroman-2-carboxylic acid(4-morpholin-4-yl-phenyl)-amide (Faster Running Isomer)

The enantiomers of theracemic-8-(4-methyl-piperazin-1-yl)-4-oxo-chroman-2-carboxylic acid(4-morpholin-4-yl-phenyl)-amide (Example 78) (100 mg, 0.22 mmol) wereseparated by the use of a chiral column (ChiralPak AD, 5 cm×50 cm, 20μ).The isomers were eluted with a gradient of 35-55% isopropanol/hexane.The faster isomer was obtained as a light yellow solid (40 mg, mp 216°C. dec.) LC/MS (M+1) m/z=451.

EXAMPLE 80

8-(4-Methyl-piperazin-1-yl)-4-oxo-chroman-2-carboxylic Acid(4-morpholin-4-yl-phenyl)-amide (Slower Running Isomer)

The enantiomers of theracemic-8-(4-methyl-piperazin-1-yl)-4-oxo-chroman-2-carboxylic acid(4-morpholin-4-yl-phenyl)-amide(100 mg, 0.22 mmol) were separated by theuse of a chiral column (ChiralPak AD, 5 cm×50 cm, 20μ). The isomers wereeluted with a gradient of 35-55% isopropanol/hexane. The slower isomerwas obtained as an off white solid (32 mg, mp 215° C. dec.) LC/MS (M+1)m/z=451.

EXAMPLE 81

4-[4-({1-[6-Fluoro-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromen-2-yl]-methanoyl}-amino)-phenyl]-piperazine-1-carboxylicAcid Ethylamide

6-Fluoro-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acid(4-piperazin-1-yl-phenyl)-amide (Example 71) (150 mg, 0.216 mmol) wasplaced in a 50 mL flask with 10 mL of CH₂Cl₂. This suspension wastreated with triethylamine (0.1 mL, 0.67 mmol) and ethylisocyanate (0.21mL, 18.7 mg, 0.26 mmol) and the reaction stirred at room temperature for18 hours. The reaction was concentrated (1 mm Hg pressure) and theconcentrate purified by chromatography on silica eluting with 1%methanol in chloroform, then concentrated (1 mm Hg pressure). Thentriturated with either to yield a yellow powder with was dried underhigh vacuum for 48 h at 50° C. (79 mg) LCMS−AP+537.4, mp=236-238° C.

EXAMPLE 82

6-Methoxy-8-(4-methyl-[1,4]diazepan-1-yl)-4-oxo-4H-chromene-2-carboxylicAcid (4-morpholin-4-yl-phenyl)-amide

Into a 100 mL round bottom flask equipped with a nitrogen inlet andmagnetic stirrer is added 327 mg (0.89 mmol, 1.0 equiv.) of6-Methoxy-8-(4-methyl-[1,4]diazepan-1-yl)-4-oxo-4H-chromene-2-carboxylicacid hydrochloride salt (Reference Example 23). This material isdissolved in 20 mL of DMF and then 189 mg (1.06 mmol, 1.2 equiv.) of4-morpholinoaniline is added. To the stirred solution is quickly addedsimultaneously added 568 mg (1.77 mmol, 2.0 equiv.) of TBTU and 239 mg(1.77 mmol, 2.0 equiv.) of HOBT. At this point 457 mg, 577 μL (25.2mmol, 4.0 equiv.) is added via syringe over 5 minutes. The reaction isallowed to stir at room temperature for 18 hrs, then is concentrated ona rotary evaporator under high vacuum in order to remove the DMF. Theresidue is triturated with methanol and the crude solids are recoveredby filtration. These residues are then purified by flash chromatographyusing a gradient of 5-10% methanol in methylene chloride as eluent. Theeluted material, which is obtained from chromatography, is concentrated,dried under high vacuum, suspended in methylene chloride, dried overK₂CO₃, concentrated, then crystallized from methanol to give the freebase of the pure product as 345 mg (79%) of a yellow solid. Mass Spec.:calc. for [C₂₇H₃₂FN₄O₅+H]⁺ Theor. m/z=393; Obs.=393.

EXAMPLE 83

6-Ethoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acid(4-morpholin-4-yl-phenyl)-amide

Into a 100 mL flask equipped with a nitrogen inlet and magnetic stirreris placed 133 mg (0.748 mmol, 1.1 equiv.) of 4-morpholinoaniline, whichis then dissolved in 20 mL of methylene chloride. To this mixture isthen added 290 mg, 367 μL (2.24 mmol, 3.3 equiv.) of ethyldiisopropylamine, followed by addition of a solution of 250 mg (0.68 mmol, 1.0equiv.) of6-ethoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carbonylchloride (Reference Example 23) which has been dissolved in 10 ml ofmethylene chloride. The reaction is allowed to stir for 4 hr, afterwhich no further formation of product was seen by LC/MS. The crudereaction was concentrated on a rotary evaporator, then triturated with10 mL of methanol. The crude solids were collected by filtration, thensubjected to flash chromatography using a gradient of from 2 to 20%methanol in methylene chloride. Recrystallization from methylenechloride and hexanes afforded 55 mg (16%) of the pure product as ayellow solid.

Mass Spec.: calc. for [C₂₇H₃₂N₄O₅+H]⁺ Theor. m/z=493; Obs. 493.

EXAMPLE 84

6-Ethoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylic acid[4-4-propionyl-piperazin-1-yl)-phenyl]-amide

This compound was prepared from 250 mg (0.68 mmol, 1.0 equiv.) of6-Ethoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carbonylchloride (Reference Example 23) and 175 mg (0.748 mmol, 1.1 equiv.) of1-[4-(4-Amino-phenyl)-piperazin-1-yl]-propan-1-one by an analogousprocedure to that used to prepare the 4-morpholino aniline derivative,to give 45 mg (12%) of the desired product as a yellow solid.

Mass Spec.: calc. for [C₃₀H₃₇N₅O₅+H]+Theor. m/z=548; Obs.=548.

EXAMPLE 85

6-Methoxy-4-oxo-8-piperazin-1-yl-4H-chromene-2-carboxylic Acid(4-morpholin-4-yl-phenyl)-amide

Into a 50 mL round bottom flask equipped with a reflux condenser,nitrogen inlet and magnetic stirrer is placed 50 mg (0.115-mmol, 1.0equiv.) of6-Methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicacid (4-morpholin-4-yl-phenyl)-amide (Example 31) and 10 mL of 1, 2dichloroethane. To this solution is then added via syringe 49 mg, 37 μL(0.345 mmol, 3.0 equiv.) of 1-chloroethyl chloroformate. A precipitateforms, indicating formation of an intermediate. The reaction is heatedto reflux for 3 days, whereupon an analysis of an aliquot by LC/MSindicates only a trace of product has formed. At this time 52 mg (0.345mmol, 3.0 equiv.) of sodium iodide are added to the refluxing reaction.LC/MS analyses then progressively show formation of demethylated productover 5 additional days. The reaction is then cooled, concentrated on arotary evaporator, then dried over K₂CO₃ as a suspension in methylenechloride containing methanol, removal of solids by filtration, followedby flash chromatography of the solution, using a gradient of 5 to 20%methanol in methylene chloride, gives 34 mg (64%) of the pure product asa reddish solid. Mass Spec.: calc. for [C₂₅H₂₉N₄O₅+H]⁺ Theor. m/z=465;Obs.=465.

EXAMPLE 86

6-Hydroxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicacid (4-morpholin-4-yl-phenyl)-amide

Into a 50 mL round bottom flask equipped with a reflux condenser,nitrogen inlet and magnetic stirrer is placed 50 mg (0.115 mmol, 1.0equiv.) of6-Methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicacid (4-morpholin-4-yl-phenyl)-amide (Example 31) and 20 mL of methylenechloride. To this solution is added 1 mL of a 1N solution of borontribromide in methylene chloride. The reaction is stirred at roomtemperature for 2.5 days at which time it is complete by LC/MS. Thereaction is concentrated on a rotary evaporator, then methanol is added.The methanol is concentrated and readded 5 times, until the BBr₃ isremoved as HBr and trimethyl borate. The solid hydrobromide saltresidue, which is obtained, is >85% pure product by LC/MS. Mass Spec.:calc. for [C₂₅H₂₈N₄O₅+H]⁺ Theor. m/z=465; obs.=465.

EXAMPLE 87 (METHOD 1)

6-Methoxy-8-(4-methyl-[1,4]diazepan-1-yl)-4-oxo-1,4-dihydro-quinoline-2-carboxylicAcid (4-morpholin-4-yl-phenyl)-amide

To a solution of6-methoxy-8-(4-methyl-[1,4]diazepan-1-yl)-4-oxo-1,4-dihydro-quinoline-2-carboxylicacid (2.10 mmol) (Reference Example 25b) and diisopropylethyl amine (1.4mL, 8.6 mmol) in 34 mL dimethylformamide was added TBTU (1.40 g, 4.36mmol) and HOBt (0.588 g, 4.35 mmol) followed by the addition of4-morpholinoaniline (0.463 g, 2.60 mmol). The resulting dark brownsolution was stirred at room temperature under nitrogen for 19 hours.The reaction was concentrated in vacuo and the resulting crude productwas taken up in methylene chloride/methanol. Filtration of the resultingmixture afforded some product as a yellow solid. The filtrates wereconcentrated and partitioned between methylene chloride and saturatedaqueous sodium bicarbonate. The organic layer was washed with saturatedsodium bicarbonate, dried (MgSO4), and concentrated under vacuum toafford a brown solid. This was suspended in methanol and filtered toafford the desired product as a yellow solid (0.714 g, 69%). ¹H NMR (300MHz, DMSO, d₆) δ 9.97 (bs, 1H, NH), 7.67 (d, 2H, J_(o)=8.8 Hz, ArH _(2′)& H _(6′)), 7.47 (bs, 1H, ArH ₅), 7.00 (s, 1H. C═CH), 6.99 (d, 2H,J_(o)=8.8 Hz, ArH _(3′) & H_(5′)), 6.71 (bs, 1H, ArH₇), 3.85 (s, 3H, OCH₃), 3.75 (t, 4H, J=4.6 Hz, OCH ₂CH₂N), 3.70 (bs, 2H, ArNCH ₂CH₂CH₂NCH₃),3.55 (bs, 2H, ArNCH₂CH₂NCH₃), 3.09 (t, 4H, J=4.6 Hz, OCH₂CH ₂N), 2.95(bs, 2H, ArNCH₂CH ₂NCH₃), 2.73 (bs, 2H, ArNCH₂CH₂CH₂NCH₃), 2.36 (s, 3H,NCHb), 2.07 (bs, 2H ArNCH₂CH ₂CH₂NCH₃); Mass Spec.: calc. for[C₂₇H₃₃N₅O₄+H]⁺ Theor. m/z=492; Obs. 492.

EXAMPLE 87 (METHOD 2)

6-Methoxy-8-(4-methyl-[1,4]diazepan-1-yl)-4-oxo-1,4-dihydro-quinoline-2-carboxylicAcid (4-morpholin-4-yl-phenyl)-amide

A solution of6-methoxy-8-(4-methyl-[1,4]diazepan-1-yl)-4-(2-trimethylsilanyl-ethoxymethoxy)-quinoline-2-carboxylicacid (4-morpholin-4-yl-phenyl)-amide (Reference. Example 27d) (0.989 g,1.59 mmol) in 20 mL methanol was poured into 300 mL 0.05 N hydrochloricacid. The clear dark yellow solution became cloudy within 5 minutes. Themixture was stirred at room temperature for 45 minutes and then adjustedto pH 7 with 10% sodium hydroxide. The resulting yellow precipitate wasisolated by filtration, washed with water, and dried under high vacuumto afford the desired product as a yellow solid (0.629 g, 80%). ¹H NMR(300 MHz, DMSO, d₆) δ 9.97 (bs, 1H,C(O)NH, 7.67 (d, 2H, J_(o)=8.8 Hz,ArH _(2′) & H _(6′)), 7.47 (bs, 1H, ArH ₅), 7.00 (s, 1H, C═CH), 6.99 (d,2H, J_(o)=8.8 Hz, ArH_(3′) & H _(5′)), 6.71 (bs, 1H, ArH ₇), 3.85 (s,3H, OCH ₃), 3.75 (t, 4H, J=4.6 Hz, OCH ₂CH₂N), 3.70 (bs, 2H, ArNCH₂CH₂CH₂NCH₃), 3.55 (bs, 2H, ArNCH ₂CH₂NCH₃), 3.09 (t, 4H, J=4.6 Hz,OCH₂CH ₂N), 2.95 (bs, 2H, ArNCH₂CH ₂NCH₃), 2.73 (bs, 2H,ArNCH₂CH₂CH_NCH₃), 2.36 (s, 3H, NCHR), 2.07 (bs, 2H ArNCH₂CH₂CH₂NCH₃);Mass Spec.: calc. for [C₂₇H₃₃N₅O₄+H]⁺ Theor. m/z=492; Obs.=492. Analysisfor C₂₇H₃₃N₅O₄. 1.0eqHCl. 0.3eqH₂O: Calculated C 60.79H 6.54 N 13.13.Found C 60.82H 6.53 N 13.17.

EXAMPLE 88

6-Methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-1,4-dihydro-quinoline-2-carboxylicAcid (4-morpholin-4-yl-phenyl)-amide

The title compound was prepared from8-bromo-6-methoxy-4-(2-trimethylsilanyl-ethoxymethoxy)-quinoline-2-carboxylicacid methyl ester (Reference Example 24c) according to the proceduresdescribed in Reference Example 25a and in Example 87 (Method 1). Ayellow solid was obtained. Mass Spec.: calc. for [C₂₆H₃₁N₅O₄+H]⁺ Theor.m/z=478; Obs. 478.

EXAMPLE 89

6-Methoxy-8-(4-methyl-piperazin-1-yl)-4-oxo-1,4-dihydro-quinoline-2-carboxylicAcid [4-(4-propionyl-piperazin-1-yl)-phenyl]-amide

The title compound was prepared from8-bromo-6-methoxy-42-trimethylsilanyl-ethoxymethoxy)-quinoline-2-carboxylicacid methyl ester (Reference Example 24c) according to the proceduresdescribed in Reference Example 25a and in Example 87 (Method 1), exceptthat the amide was formed from1-[4-(4-amino-phenyl)-piperazin-1-yl]-propan-1-one. A yellow solid wasobtained. Mass Spec.: calc. for [C₂₉H₃₆N₆O₄+H]⁺ Theor. m/z=533; Obs.533.

EXAMPLE 90

6-Fluoro-8-(4-methyl-piperazin-1-yl)-4-oxo-1,4-dihydro-quinoline-2-carboxylicAcid (4-morpholin-4-yl-phenyl)-amide

The title compound was prepared from6-Fluoro-8-(4-methyl-piperazin-1-yl)-4-oxo-1,4-dihydro-quinoline-2-carboxylicacid hydrochloride salt (Reference Example 26) using the proceduredescribed in Example 87 (Method 1). After chromatography, it is thencrystallized from methanol to give the pure product as 150 mg (55%) of ayellow solid. Mass Spec.: calc. for [C₂₅H₂₈FN₅O₃+H]⁺ Theor. m/z=466;Obs.=466.

EXAMPLE 91

6-Fluoro-8-(4-methyl-piperazin-1-yl)-4-oxo-1,4-dihydro-quinoline-2-carboxylicacid [4-(4-propionyl-piperazin-1-yl)-phenyl]-amide

The title compound was prepared from6-Fluoro-8-(4-methyl-piperazin-1-yl)-4-oxo-1,4-dihydro-quinoline-2-carboxylicacid hydrochloride salt (200 mg, 0.59 mmol) (Reference Example 26) usingthe procedure described in Example 87 (Method 1). 31% yield. Mass Spec.:calc. for [C₂₈H₃₃FN₆O₃+H]⁺ Theor. m/z=521; Obs.=521.

EXAMPLE 92

8-[(2-Dimethylamino-ethyl)-methyl-amino]-6-methoxy-4-oxo-1,4-dihydro-quinoline-2-carboxylicacid (4-morpholin-4-yl-phenyl)-amide

The title compound was prepared from8-bromo-6-methoxy-4-(2-trimethylsilanyl-ethoxymethoxy)-quinoline-2-carboxylicacid methyl ester (Reference Example 24c) according to the proceduresdescribed in Reference Example 25a and in Example 87 (Method 2), usingN,N,N′-trimethyl ethylenediamine for the Pd catalysed coupling. A yellowsolid was obtained. Mass Spec.: calc. for [C₂₆H₃₃N₅O₄+H]⁺ Theor.m/z=480; Obs.=480.

EXAMPLE 93

8-[(3-Dimethylamino-propyl)-methyl-amino]-6-methoxy-4-oxo-1,4-dihydro-quinoline-2-carboxylicacid (4-morpholin-4-yl-phenyl)-amide

The title compound was prepared from8-bromo-6-methoxy-4-(2-trimethylsilanyl-ethoxymethoxy)-quinoline-2-carboxylicacid methyl ester (Reference Example 24c) according to the proceduresdescribed in Reference Example 25a and in Example 87 (Method 2), usingN,N,N′-trimethyl-1,3-propanediamine for the Pd catalysed coupling. Ayellow solid was obtained. Mass Spec.: calc. for [C₂₇H₃₅N₅O₄+H]⁺ Theor.m/z=494; Obs.=494.

EXAMPLE 94

8-((3R)-(+)-3-Dimethylamino-pyrrolidino-1-yl)-6-methoxy-4-oxo-1,4-dihydro-quinoline-2-carboxylicAcid (4-morpholin-4-yl-phenyl)-amide

The title compound was prepared from8-bromo-6-methoxy-4-(2-trimethylsilanyl-ethoxymethoxy)-quinoline-2-carboxylicacid methyl (Reference Example 24c) according to the proceduresdescribed in Reference Example 25a and in Example 87 (Method 2), using(3R)-(+)-3-(dimethylamino)pyrrolidine for the Pd catalysed coupling. Ayellow solid was obtained. Mass Spec.: calc. for [C₂₇H₃₃N₅O₄+H]⁺ Theor.m/z=492; Obs.=492.

EXAMPLE 95

8-((3S)-(−)-3-Dimethylamino-pyrrolidin1-yl)-6-methoxy-4-oxo-1,4-dihydro-quinoline-2-carboxylic Acid(4-morpholin-4-yl-phenyl)-amide

The title compound was prepared from8-bromo-6-methoxy-4-(2-trimethylsilanyl-ethoxymethoxy)-quinoline-2-carboxylicacid methyl ester (Reference Example 24c) according to the proceduresdescribed in Reference Example 25a and in Example 87 (Method 2), using(3S)-(−)-3-(dimethylamino)pyrrolidine for the Pd catalysed coupling. Ayellow solid was obtained. Mass Spec.: calc. for [C₂₇H₃₃N₅O₄+H]⁺ Theor.m/z=492; Obs.=492.

EXAMPLE 96

6-Methoxy-8-[methyl-(1-methyl-pyrrolidin-3-yl)-amino]4-oxo-1,4-dihydro-quinoline-2-carboxylicAcid (4-morpholin-4-yl-phenyl)-amide

The title compound was prepared from8-bromo-6-methoxy-4-(2-trimethylsilanyl-ethoxymethoxy)-quinoline-2-carboxylicacid methyl ester (Reference Example 24c) according to the proceduresdescribed in Reference Example 25a and in Example 87 (Method 2), usingN,N′-dimethyl-3-aminopyrrolidine for the Pd catalysed coupling. A yellowsolid was obtained. Mass Spec.: calc. for [C₂₇H₃₃N₅O₄+H)⁺ Theor. m/z492; Obs.=492.

EXAMPLE 97

8-[Ethyl-(1-ethyl-pyrrolidin-3-yl)-amino]-6]-6-methoxy-4-oxo-1,4-dihydro-quinoline-2-carboxylicAcid (4-morpholin-4-yl-phenyl)-amide

The title compound was prepared from8-bromo-6-methoxy-4-(2-trimethylsilanyl-ethoxymethoxy)-quinoline-2-carboxylicacid methyl ester (Reference Example 24c) according to the proceduresdescribed in Reference Example 25a and in Example 87 (Method 2), using3-diethylaminopyrrolidine for the Pd catalyzed coupling. A yellow solidwas obtained. Mass Spec.: calc. for [C₂₉H₃₇N₅O₄+H]⁺ Theor. m/z=520;Obs.=520.

EXAMPLE 98

4-Dimethylamino-6-methoxy-8-(4-methyl-piperazin-1-yl)-quinoline-2-carboxylicAcid (4-morpholin-4-yl-phenyl)-amide

To a suspension of8-bromo-4-dimethylamino-6-methoxy-quinoline-2-carboxylic acid(4-morpholin-4-yl-phenyl)-amide (Reference Example 28b) (139.9 mg, 0.288mmol), N-methylpiperazine (48 mL, 0.43 mmol), and 4 Å sieves in 15 mLanhydrous toluene was added Pd₂ (dba)₂ (15.3 mg, 16.7 μmol), BINAP (63.0mg, 0.101 mmol) and cesium carbonate (0.436 g, 1.345 mmol). Theresulting wine colored mixture was heated at reflux under nitrogen for20 hours. The reaction mixture was cooled to room temperature andconcentrated. The crude mixture was purified by flash chromatography onsilica gel using a gradient of 100:0 to 95:5 methylene chloride:methanolto afford the desired product as a yellow solid (96.9 mg, 67%). ¹H NMR(300 MHz, DMSO, d₆) δ 10.06 (s, 1H, C(O)NH), 7.69 (d, 2H, J_(o)=9.0 Hz,ArH _(2′) & H _(6′)), 7.58 (s, 1H, ArH ₃), 7.58 (d, 2H, J_(o)=9.0 Hz,ArH _(3′) & H _(5′)), 6.95 (d, 1H, J_(m)=2.7 Hz, ArH ₅), 6.76 (d, 1H,J_(m)=2.7 Hz, ArH ₇), 3.90 (s, 3H, OCH ₃), 3.75 (t, 4H, J=4.8 Hz, OCH₂CH₂N), 3.37 (bs, 4H, ArNCH ₂CH₂N), 3.10 (t, 4H, J=4.8 Hz, OCH₂CH ₂N),3.01 (s, 6H, N(CH ₃)₂), 2.71 (bs, 4H, ArNCH₂CH ₂N), 2.35 (s, 3H, R₂NCH₃); Mass Spec.: calc. for [C₂₈H₃₆N₆O₃+H]⁺ Theor. m/z=505; Obs.=505.5.

EXAMPLE 99

6-Methoxy-4-methylamino-8-(4-methyl-piperazin-1-yl)-quinoline-2-carboxylicAcid (4-morpholin-4-yl-phenyl)-amide

The title compound was prepared from8-bromo-6-methoxy-4-oxo-1,4-dihydro-quinoline-2-carboxylic acid(Reference Example 27b) according to the procedure described for Example98 using N-methyl amine to prepare8-bromo-4-methylamino-6-methoxy-quinoline-2-carboxylic acid(4-morpholin-4-yl-phenyl)-amide. A glassy orange solid was obtained.Mass Spec.: calc. for [C₂₇H₃₄N₆O₃+H]⁺ Theor. m/z=491; Obs.=491.5.

EXAMPLE 100

6-Fluoro-4-methoxy-8-(4-methyl-piperazin-1-yl)-quinoline-2-carboxylicAcid (4-morpholin-4-yl-phenyl)-amide

Into a 250 mL round bottom flask equipped with a nitrogen inlet andmagnetic stirrer is added 2.01 g (6.3 mmol, 1.0 equiv.) of6-Fluoro-4-methoxy-8-(4-methyl-piperazin-1-yl)-quinoline-2-carboxylicacid hydrochloride salt. This material is dissolved in 20 mL of DMF andthen 1.35 g (7.56 mmol, 1.2 equiv.) of 4-morpholinoaniline is added. Tothe stirred solution is quickly added simultaneously added 4.05g (12.6mmol, 2.0 equiv.) of TBTU(2-(1H-benzotriazole-1-yl)-1,1,3,3tetramethyluroniumtetrafluoroborate)and 1.7 g (12.6 mmol, 2.0 equiv.) of HOBT (1-hydroxybenzotriaolehydrate). At this point 3.25 g, 4.11 mL (25.2 mmol, 4.0 equiv.) is addedvia syringe over 5 minutes. The reaction is allowed to stir at roomtemperature for 18 hrs, then is concentrated on a rotary evaporatorunder high vacuum in order to remove the DMF. The residue is trituratedwith methanol and the crude solids are recovered by filtration. Thematerial is then dissolved in methylene chloride and extracted with 10%sodium bicarbonate solution. The organic layer is dried and thenconcentrated. These residues are then purified by flash chromatographyusing a gradient of 5-10% methanol in methylene chloride as eluent. Thematerial which is obtained from chromatography, is then crystallizedfrom methanol to give the pure product as 2.83g (93%) of a yellow solid.Mass Spec.: calc. for [C₂₆H₃₀FN₅O₃+H]⁺ Theor. m/z=480; Obs.=480.

EXAMPLE 101 6-Fluoro-4-oxo-8-piperazin-1-yl-4H-chromene-2-carboxylicacid (4-morpholin-4-yl-phenyl)-amide: Made According to the GeneralMethod of Howarth et. al. Tetrahedron, 1998, 54, 10899-10914

Dry 6-flouro-8-(4-methyl-piperazin-1-yl)-4-oxo-4H-chromene-2-carboxylicacid [4-(4-propionyl-piperazin-1-yl)-phenyl]-amide (Example 72)(1 g 1.9mmol) was added to 100 mL of rigorously dried 1,2-dichloroethane in aflask under N₂ atmosphere and magnetic stirring. The mixture was cooledto 0° C. and freshly distilled 1-chloroethyl chloroformate (650 ul, 858mg, 6 mmol, 3 eq) was added drop wise. The reaction was then heatedunder reflux for 5 hours at which time LC/MS revealed completeconsumption of starting material. NaI (1 g, 1 eq) was added and heatingcontinued for 2 days more. The reaction was then allowed to cool andfiltered and evaporated to dryness under reduced pressure. MeOH (100 mL)was added and heated to reflux for 4h, filtered hot and evaporated todryness. The product was isolated by chromatography using silica gel andCHCl3/5% MeOH as an eluent. This gave 700 mg of the product HCl salt asa yellow solid. LCMS−m/z=508.

1. A compound represented by the formula (I):

wherein R¹ is, at each position, independently represented by hydrogen,optionally substituted alkyl, optionally substituted cycloalkyl,methoxy, thiomethoxy, —NHA, —NA₂, —NHC(═O)A, aminocarbonyl, —C(═O)NHA,—C(═O)NA₂, halogen, hydroxy, —OA, cyano or aryl; A is optionallysubstituted alkyl, optionally substituted cycloalkyl, optionallysubstituted alkenyl or optionally substituted alkynyl; R² is representedby (i), (ii), (iii), or (iv) below:

R³ is —H, optionally substituted C₁₋₆alkyl, optionally substitutedC₂₋₆alkenyl, optionally substituted C₂₋₆alkynyl, optionally substitutedC₃₋₆cycloalkyl or AOH; n is 2, 3 or 4; P is a heterocyclic ring; R⁴ is—H or optionally substituted C₁₋₄alkyl; R⁵ is ═O, ═NR⁴ or ═S; R⁶ is —Hor methyl; Y is —C(═O)NH—, —C(═O)NA-, —C(═O)N(A)-, —NHC(═O)—, —C(═S)NH—,—CH₂NH—, —C(═O)CH₂—, —CH₂C(═O)—, —C(═O)-piperazine-, —C(═O)R⁸—,—NAC(═O)—, —C(═S)N(A)-, —CH₂N(A)-, —N(A)CH₂— or a 5-memberedheterocyclic. R⁷ is a monocyclic or bicyclic aromatic ring or aheterocycle, optionally substituted by one or more substituents selectedfrom R⁸-R⁹ and R¹⁰; wherein R is connected to Y either by a single bondor by a ring fusion; R⁸ is —CH₂—, —C(═O)—, —SO₂—, —SO₂NH—, —C(═O)NH—,—O—, —S—, —S(═O)—, a five-membered heterocyclic connected to R⁷ by aring fusion or a single bond as tether; R⁹ is morpholine optionallysubstituted with at least one substituent selected from A,thiomorpholine, piperazin-R¹¹, optionally substituted aryl, optionallysubstituted heterocycle, or —C(═O)CA; R¹⁰ is optionally substitutedalkyl, optionally substituted cycloalkyl, hydroxy, aryl, cyano, halogen,—C(═O)NH₂—, methylthio, —NHA, —NA₂, —NHC(═O)A, —C(═O)NHA, —C(═O)NA₂, orOA; R¹¹ is —H, alkyl, AOH, —SO₂A, —SO₂NH₂, —SO₂NHA, —SO₂NA₂, —SO₂NHAR⁹,—C(═O)R⁹, -alkylR⁹, C(═O)A, C(═O)NH₂, C(═O)NHA, C(═O)NA₂, or —C(═O)OA.2. The composition as recited in claim 1 wherein the R¹ is, at eachposition, independently represented by hydrogen, alkyl, cycloalkyl,methoxy, thiomethoxy, —NHA, —NA₂, —NHC(═O)A, aminocarbonyl, —C(═O)NHA,—C(═O)NA₂, halogen, hydroxy, —OA, cyano, or aryl wherein alkyl andcycloalkyl are optionally substituted with halogen, nitro, cyano,hydroxy, trifluoromethyl, amino, carboxy, carboxamindo, amidino,carbamoyl, mercapto, sulfamoyl, C₁₋₄ alkyl, C₂₋₄ alkenyl, C₂₋₄ alkynyl,C₃₋₆ cycloalkyl, C₃₋₆ cycloalkenyl, C₁₋₄ alkoxy, C₁₋₄ alkanoyl, C₁₋₄alkanoyloxy, N-(C₁₋₄ alkyl), N(C₁₋₄ alkyl)₂, C₁₋₄ alkanoylamino, (C₁₋₄alkanoyl)₂amino, N-(C₁₋₄alkyl)carbamoyl, N,N-(C₁₋₄)₂carbamoyl, (C₁₋₄alkyl)S, (C₁₋₄ alkyl)S(═O), (C₁₋₄ alkyl)SO₂, (C₁₄) alkoxycarbonyl,N-(C₁₋₄ alkyl)sulfamoyl, N,N-C₁₋₄ alkyl)sulfamoyl, C₁₋₄alkylsolfonylamino or heterocyclic.
 3. The composition as recited inclaim 1 wherein A represents an alkyl optionally substituted withhalogen, nitro, cyano, hydroxy, trifluoromethyl, amino, carboxy,carboxamindo, amidino, carbamoyl, mercapto, sulfamoyl, C₁₋₄ alkyl, C₂₋₄alkenyl, C₂₋₄ alkynyl, C₃₋₆ cycloalkyl, C₃₋₆ cycloalkenyl, C₁₋₄ alkoxy,C₁₋₄ alkanoyl, C₁₋₄ alkanoyloxy, N-(C₁₋₄ alkyl), N(C₁₋₄ alkyl)₂, C₁₋₄alkanoylamino, (C₁₋₄ alkanoyl)₂amino, N-(C₁₋₄ alkyl)-carbamoyl,N,N-(C₁₄)₂carbamoyl, (C₁₋₄alkyl)S, (C₁₋₄ alkyl)S(═O), (C₁₋₄alkyl)SO₂,(C₁₄) alkoxycarbonyl, N-(C₁₋₄ alkyl)sulfamoyl, N,N-C₁₋₄ alkyl)sulfamoyl,C₁₋₄ alkylsolfonylamino, or heterocyclyl.
 4. The composition as recitedin claim 3 wherein R³ is —H, C₁₋₆alkyl, C₂₋₆alkenyl, C₂₋₆alkynyl,C₃₋₆cycloalkyl or AOH; wherein the C₁₋₆alkyl, C₂₋₆alkenyl, C₂₋₆alkynyl,C₃₋₆cycloalkyl and A are optionally substituted with halogen, nitro,cyano, hydroxy, trifluoromethyl, amino, carboxy, carboxamindo, amidino,carbamoyl, mercapto, sulfamoyl, C₁₋₄ alkyl, C₂₋₄alkenyl, C₂₋₄ alkynyl,C₃₋₆ cycloalkyl, C₃₋₆ cycloalkenyl, C₁₋₄ alkoxy, C₁₋₄ alkanoyl, C₁₋₄alkanoyloxy, N-(C₁₋₄ alkyl), N(C₁₋₄ alkyl)₂, C₁₋₄ alkanoylamino, (C₁₋₄alkanoyl)₂amino, N-(C₁₋₄alkyl)carbamoyl, N,N-(C₁₋₄)₂carbamoyl, (C₁₋₄alkyl)S, (C₁₋₄ alkyl)S(═O), (C₁₋₄ alkyl)SO₂, (C₁₋₄) alkoxycarbonyl,N-(C₁₋₄ alkyl)sulfamoyl, N,N-C₁₋₄ alkyl)sulfamoyl, C₁₋₄alkylsolfonylamino, or heterocyclic.
 5. The composition as recited inclaim 1 wherein R⁷ represents a monocyclic or bicyclic aromatic ringoptionally incorporating at least one heteroatom selected from N, O andS.
 6. The composition as recited in claim 5 wherein the aromatic ringoptionally incorporating a heteroatom comprises phenyl, 1- and2-naphthyl, 2-, 3- and 4-pyridyl, 2- and 3-thienyl, 2- and 3-furyl,quinolyl, isoquinolyl, indolyl, benzothienyl, benzofuryl, 1-, 2- and3-pyrrolyl, imidazolyl, thiazolyl, oxazolyl, pyrazolyl, isothiazolyl,isoxazolyl, 1,2,3-triazolyl, 1,2,3-thiadiazolyl, 1,2,3-oxadiazolyl,1,2,4-triazolyl, 1,2,4-thiadiazolyl, 1,2,4-oxadiazolyl, 1,3,4-triazolyl,1,3,4-thiadiazolyl, 1,3,4 oxadiazolyl, benzimidazolyl, benzthiazolyl,benzoxazolyl or triazinyl.
 7. The composition as recited in claim 6wherein R⁷ is represented by the formula (v):

wherein R⁷ is optionally substituted by at least one substituentselected from R⁸-R⁹ and R¹⁰ wherein R represents a single bond, —C(═O)—,—CH₂—, —O—, —S—, —SO₂—, or —S(═O)— as tether, or a five-memberedheterocycle connected to R⁷ by a ring fusion, and R⁹ is morpholineoptionally substituted with at least one substituent selected from A,thiomorpholine, a 5-membered heterocycle, —C(═O)A, an aryl or ahetercyclic; wherein the aryl or heterocyclic is optionally substitutedwith halogen, nitro, cyano, hydroxy, trifluoromethyl, amino, carboxy,carboxamindo, amidino, carbamoyl, mercapto, sulfamoyl, C₁₋₄ alkyl, C₂₋₄alkenyl, C₂₋₄ alkynyl, C₃₋₆ cycloalkyl, C₃₋₆ cycloalkenyl, C₁₋₄ alkoxy,C₁₋₄ alkanoyl, C₁₋₄ alkanoyloxy, NH-(C₁₋₄ alkyl), N(C₁₋₄ alkyl)₂, C₁₋₄alkanoylamino, (C₁₋₄ alkanoyl)₂amino, N—(C₁₋₄alkyl)carbamoyl,N,N-(C₁₋₄)₂carbamoyl, (C₁₋₄ alkyl)S—, (C₁₋₄ alkyl)S(═O)—, (C₁₋₄alkyl)SO₂—, (C₁₋₄)alkoxycarbonyl, N-(C₁₋₄ alkyl)sulfamoyl, N,N-C₁₋₄alkyl)sulfamoyl, C₁₋₄ alkylsolfonylamino or heterocyclyl.
 8. Thecomposition as recited in claim 7 wherein R¹⁰ is alkyl, hydroxy, cyano,OA or halogen.
 9. The composition as recited in claim 8 wherein R¹⁰ iscyano, hydroxy, methoxy, ethoxy or halogen.
 10. The composition asrecited in claim 1 wherein R⁷ is represented by the Formula (vi):

wherein R⁸ is a single bond as tether and R⁹ is methoxy, cyano, afive-membered heterocycle or a compound represented by the Formula(vii):


11. The composition as recited in claim 10 wherein R¹¹ is —H, —SO₂CH₃,—SO₂CH₂CH₃, —SO₂-n-C₃H₇, —SO₂-1-C₃H₇, —SO₂-nC₄H₁₀, —SO₂-t-C₄H₁₀,—SO₂NH₂, SO₂N(CH₃)₂, —C(═O)NH₂, —C(═O)NH-cyclohexyl, —C(═O)-cyclopentyl,—C(═O)-pyrrolidine, —C(═O)N(CH₃)₂, —C(═O)-morpholine, —C(═O)CH₃,—C(═O)CH₂CH₃, —C(═O)-n-C₃H₇, —C(═O)-1-C₃H₇, —C(═O)-n-C₄H₁₀,—C(═O)-1-C₄Hio, —C(═O)-t-C₄H₁₀, CH₃OH, SO₂CH(CH₃)₂, SO₂NHCH₂CH(CH₃)₂,—CH₂CH₂OH, —C(═O)CH₂CH₂OH, —C(═O)NHCH₂CH₃ or C(═O)OC₄H₁₀.
 12. Thecompound as recited in claim 1 wherein Y represents a five-memberedheterocyclic ring comprising one or more heteroatoms selected from S, Nand O.
 13. The compound as recited in claim 12 wherein Y is pyrrole,thiophene, furan, imidazole, thiazole, oxazole, pyrazole, isothiazole,isoxazole, 1,2,3-triazole, 1,2,3-thiadiazole, 1,2,3-oxadiazole,1,2,4-triazole, 1,2,4-thiadiazole, 1,2,4-oxadiazole, 1,3,4-triazole,1,3,4-thiadiazole or 1,3,4-oxadiazole.
 14. The compound as recited inclaim 12 wherein Y is —C(═O)NH—, —C(═O)N(CH₃)—, —NHC(═O)— or—C(═O)-piperazine-.
 15. A compound represented by the formula (I):

wherein R¹ is, at each position, independently represented by hydrogen,alkyl, optionally substituted cycloalkyl, methoxy, thiomethoxy, —NHA,—  NA₂, —NHCOA, aminocarbonyl, —CONHA, —CONA₂, halogen, hydroxy, —OA,cyano or aryl; A is alkyl, cycloalkyl, alkenyl or alkynyl; R² isrepresented by (i), (ii), (iii), or (iv) below:

R³ is —H, C₁₋₆alkyl, C₂₋₆alkenyl, C₂₋₆alkynyl, C₃₋₆cycloalkyl or AOH; Pis a heterocyclic ring; n is 2 or 3; R⁵ is =O; R⁶ is —H or methyl; Y is—C(═O)NH—, —C(═O)NA-, —C(═O)N(A)-, —NHC(═O)—, —C(═S)NH—, —CH₂NH—,—C(═O)CH₂—, —CH₂C(═O)—, —C(═O)-piperazine-, —C(═O)R⁸—, —NAC(═O)—,—C(═S)N(A)-, —CH₂N(A)-, —N(A)CH₂— or a 5-membered heterocyclic; R⁷ is amonocyclic or bicyclic aromatic ring or a heterocycle, optionallysubstituted by one or more substituents selected from R⁸-R⁹ and R¹⁰;wherein R⁷ is connected to Y either by a single bond or by a ringfusion; R⁸ is —CH₂—, —C(═O)—, —SO₂—, —SO₂NH—, —C(═O)NH—, —O—, —S—,—S(═O)—, a five-membered heterocyclic connected to R⁷ by a ring fusionor single bond as tether; R⁹ is morpholine optionally substituted withat least one substituent selected from A, thiomorpholine, piperazin-R¹¹,optionally substituted aryl, optionally substituted heterocyclic, or—C(═O)CA; R¹⁰ is optionally substituted alkyl, optionally substitutedcycloalkyl, hydroxy, aryl, cyano, halogen, —C(═O)NH₂—, methylthio, —NHA,—NA₂, —NHC(═O)A, —C(═O)NHA, —C(═O)NA₂ or OA; R¹¹ is —H, alkyl, AOH,—SO₂A, —SO₂NH₂, —SO₂NHA, —SO₂NA₂, —SO₂NHAR⁹, —C(═O)R⁹, -alkylR⁹, C(═O)A,C(═O)NH₂, C(═O)NHA, C(═O)NA₂ or —C(═O)OA.
 16. A compound as recited inclaim 15 wherein R¹ is, at each position, independently represented byhydrogen or an alkyl having 1-6 carbon atoms.
 17. A compound as recitedin claim 15 wherein R¹ is, at each position, independently representedby a cycloalklyl having 3-6 carbon atoms.
 18. A compound as recited inclaim 15 wherein R¹ is, at each position, independently represented byhydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl,tert-butyl, cyclopentyl or cyclohexyl.
 19. A compound as recited inclaim 15 wherein R¹ is, at each position, independently represented byOA, wherein A represents alkyl.
 20. A compound as recited in claim 15wherein R¹ is, at each position, independently represented by halogen.21. A compound as recited in claim 20 wherein R¹ is, at each position,independently represented by bromine, chlorine or fluorine.
 22. Acompound as recited in claim 15 wherein R¹ is, at each position,independently represented by hydrogen.
 23. A compound as recited in anyone of claims 15-22 wherein R³ is —H or C₁₋₆ alkyl.
 24. A compound asrecited in any one of claims 15-22 wherein R⁷ is represented by theFormula (v):

wherein R⁷ is optionally substituted by at least one substituentselected from R⁸-R⁹ and R¹⁰ wherein R⁸ is a single bond as tether or afive-membered heterocycle connected to R⁷ by a ring fusion and R⁹ ismorpholine, thiomorpholine or C(═O)A.
 25. A compound as recited in claim15 wherein R¹⁰ is at least one substituent selected from alkyl,cycloalkyl, OA, halogen, and cyano.
 26. A compound as recited in claim24 wherein R¹⁰ is cyano, hydroxy, methoxy, ethoxy, chlorine, bromine orfluorine.
 27. A compound as recited claim 25 wherein R⁷ is representedby the formula (vi):

wherein R⁸ is a single bond as tether and R⁹ is methoxy, cyano or acompound represented by the Formula (vii):

wherein R¹¹ is —SO₂CH₃, —SO₂CH₂CH₃, —SO₂-n-C₃H₇, —SO₂-1-C₃H₇,—SO₂-n-C₄H₀, —SO₂-1-C₄H₁₀, —SO₂-t-C₄H₁₀, —SO₂NH₂, —SO₂N(CH₃)₂,—C(═O)NH₂, —C(═O)NCH₂CH₃, C(═O)NH-cyclo-C₆H₁₂, C(═O)-cyclo-C₅H₁₀,—C(═O)-pyrrolidone, —C(═O)N(CH₃)₂, —C(═O)-morpholine, —C(═O)CH₃,—C(═O)CH₂CH₃, —C(═O)-n-C₃H₇, —C(═O)-1-C₃H₇, C(═O)CH₂CH₂OH,C(═O)-n-C₄H₁₀, —C(═O)-1-C₄H₁₀, —C(═O)-t-C₄H₁₀, —CH₃OH, CH₂CH₂OH or—C(═O)OC₄H₁₀.
 28. A compounds as recited in claim 27 wherein Y is afive-membered heterocyclic ring comprising one or more heteroatomsselected from S, N and O.
 29. A compound as recited in claim 27 whereinY is furan, diazol, oxadiazole, pyrrole, thiophene, furan, imidazole,thiazole, oxazole, pyrazole, isothiazole, isoxazole, 1,2,3-triazole,1,2,3-thiadiazole, 1,2,3-oxadiazole, 1,2,4-triazole, 1,2,4-thiadiazole,1,2,4-oxadiazole, 1,3,4-triazole, 1,3,4-thiadiazole or 1,3,4-oxadiazole.30. A compound as recited in claim 29 wherein Y is —C(═O)NH—,—C(═O)N(CH₃)—, —NHC(═O)— or —C(═O)-piperazine-.
 31. A compoundrepresented by the formula (I):

wherein R¹ is, at each position, independently represented by hydrogen,C₁₋₆ alkyl, halogen, hydroxy, —OA or cyano; A is alkyl, cycloalkyl,alkenyl or alkynyl; R² is:

R³ is —H or C₁₋₆alkyl; n is 2; R⁵ is ═O; R⁶ is —H or methyl; Y is—C(═O)NH or —C(═O)-piperazine-; R⁷ is phenyl optionally substituted byone or more substituents selected from R⁸-R⁹ or R¹⁰; R⁸ is afive-membered heterocycle connected to R⁷ by a ring fusion or a singlebond as tether; R⁹ is morpholine, thiomorpholine, —C(═O)A orpiperazin-R¹¹; R¹⁰ is alkyl, hydroxy, cyano, halogen or OA; R¹¹ is —H,alkyl, AOH, —SO₂A, —SO₂NH₂, —SO₂NHA, —SO₂NA₂, —SO₂NHAR⁹, —C(═O)R⁹,-alkylR⁹, —C(═O)A, —C(═O)NH₂, —C(═O)NHA, —C(═O)NA₂ or —C(═O)OA; or apharmaceutically acceptable salt of said compound.
 32. A compound asrecited in claim 31 wherein R¹ is, at each position, independentlyrepresented by —H, C₁-C₆ alkyl, C₃₋₆cycloalkyl, halogen or OA.
 33. Acompound as recited in claim 32 wherein R¹ is, at each position,independently represented by fluorine, chorine, bromine, methoxy, ethoxyor methyl.
 34. A compound as recited in claim 33 wherein R³ is methyl.35. A compound as recited in claim 34 wherein Y is —C(═O)NH—.
 36. Acompound as recited in claim 35 wherein R⁷ is phenyl optionallysubstituted by one or more substituents selected from R⁸-R⁹ and R¹⁰wherein R⁸ is a five-membered heterocyclic connected to R⁷ by a ringfusion or a single bond as tether and R⁹ is morpholine optionallysubstituted on carbon with at least one substituent selected from A,piperazine-R¹¹ and thiomorpholine.
 37. A compound as recited in 36wherein R⁹ is morpholine.
 38. A compound as recited in claim 37 whereinR¹⁰ is cyano, OA, OH or halogen.
 39. A compound as recited in claim 38wherein R¹⁰ is cyano, methoxy, ethoxy, fluorine, chlorine, bromine orhydroxyl.
 40. A compound of claim 36 wherein R⁹ is piperazine-R¹¹,wherein R¹ is —SO₂CH₃, —SO₂CH₂CH₃, —SO₂-n-C₃H₇, —SO₂-i-C₃H₇,—SO₂-n-C₄H₁₀, —SO₂-i-C₄H₁₀, —SO₂-t-C₄H₁₀, —SO₂NH₂, —SO₂N(CH₃)₂,—C(═O)NH₂, —C(═O)NCH₂CH₃, C(═O)NH-cyclo-C₆H₁₂, —C(═O)-cyclo-C₅H₁₀,—C(═O)-pyrrolidone, —C(═O)N(CH₃)₂, —C(═O)-morpholine, —C(═O)CH₃,—C(═O)CH₂CH₃, —C(═O)-n-C₃H₇, —C(═O)-i-C₃H₇, C(═O)CH₂CH₂OH,C(═O)-n-C₄H₁₀, —C(═O)-i-C₄H₁₀, —C(═O)-t-C₄H₁₀, —CH₃OH, CH₂CH₂OH or—C(═O)OC₄H₁₀.
 41. A compound according to claim 1, 15 or 31 for use inthe treatment of depression, generalized anxiety, eating disorders,dementia, panic disorder, sleep disorders, gastrointestinal disorders,motor disorders, endocrine disorders, vasospasm and sexual dysfunctionof an animal in need of such therapy.
 42. A method of treatment of ahuman or animal suffering from depression, generalized anxiety, eatingdisorders, dementia, panic disorder, sleep disorders, gastrointestinaldisorders, motor disorders, endocrine disorders, vasospasm and sexualdysfunction administering to such animal an effective amount of acompound of Formula I according to claim 1, 15 or 31 or apharmaceutically acceptable salt of said compound.
 43. (canceled)